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  • 89
    STEMCELL Technologies Inc mscs mscs
    <t>CTLA-4</t> affected the tumourigenic capacity of melanoma cells. C57BL/6 mice were inoculated with (A) B16-F0 cells and (B) B16-F1 cells for 7 days, separately. C57BL/6 mice were then administered with CTLA-4 inhibitor. The tumour size of the melanoma was monitored. The residual ALDH + <t>MSCs</t> within the tumour were analysed by flow cytometry in a (C) B16-F0 mouse model and (D) B16-F1 mouse model. **P
    Mscs Mscs, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mscs mscs - by Bioz Stars, 2020-08
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    92
    Biological Industries Inc mscs
    <t>CTLA-4</t> affected the tumourigenic capacity of melanoma cells. C57BL/6 mice were inoculated with (A) B16-F0 cells and (B) B16-F1 cells for 7 days, separately. C57BL/6 mice were then administered with CTLA-4 inhibitor. The tumour size of the melanoma was monitored. The residual ALDH + <t>MSCs</t> within the tumour were analysed by flow cytometry in a (C) B16-F0 mouse model and (D) B16-F1 mouse model. **P
    Mscs, supplied by Biological Industries Inc, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Cyagen Biosciences mscs
    Infused mesenchymal stem cells <t>(MSCs)</t> persist in spleen after ischemia/reperfusion injury (IRI). Intravenously delivered <t>RFP-labeled</t> MSCs were not detected in ischemic kidney at any time point ( a ), but persisted in spleen during the whole process, at least for 120 h after reperfusion ( b ). RFP-labeled MSCs (red) were detected by laser confocal microscopy. Nuclei were stained with DAPI (blue). Original magnification, × 600, n =3. ( c ) Intravenously delivered DiR-labeled MSCs (yellow and red) persisted in the spleen at least for 120 h after reperfusion. Signals were detected by IVIS in vivo imaging, n =3.
    Mscs, supplied by Cyagen Biosciences, used in various techniques. Bioz Stars score: 92/100, based on 181 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 181 article reviews
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    mscs - by Bioz Stars, 2020-08
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    mscs  (Lonza)
    94
    Lonza mscs
    MIIA is more assembled on stiff matrix than soft, and the phosphomimetic S1943D is more soluble and mobile than WT or S1943A. (A, i) <t>MSCs</t> <t>transfected</t> with GFP-MIIA or GFP-MIIB on stiff matrix were photobleached along actin bundles and allowed to recover. MIIA recovered faster and thus was more mobile than MIIB. (ii) MSCs expressing GFP-MIIA on either soft 1-kPa matrix or stiff 34-kPa matrix were similarly analyzed, and MIIA in cells on soft matrix recovered more quickly. (iii) Phosphomimetic GFP-MIIA S1943D mutant recovered faster than GFP-MIIA S1943A, which was less than or equal to the behavior of GFP-MIIA on stiff matrix. Data are means ± SEM for at least five cells. (B) Fraction of insoluble MIIA is quantified by immunofluorescence of cell ghosts that were derived from Triton X-100 extraction of cells on matrices for 4 min (±SEM; n = 3). Western blotting of the insoluble fraction of the cells on matrices was also performed. Representative blot for MIIA is shown, normalized to total protein. (C) MSCs transfected with either GFP only, GFP-MIIA (WT), S1943D, or S1943A mutants were Triton X-100 extracted, and the extracted amount of MIIA (soluble) was compared with the insoluble fraction. Immunoblots show anti-MIIA blotting with β-actin as a loading control. When the amount of MIIA is normalized to β-actin, the only significant difference in the insoluble/soluble MIIA ratio is between S1943D and S1943A (P ≤ 0.05; ±SEM; n = 3). Endog., endogenous.
    Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 94/100, based on 1176 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mscs - by Bioz Stars, 2020-08
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    92
    MEDIPOST mscs
    MIIA is more assembled on stiff matrix than soft, and the phosphomimetic S1943D is more soluble and mobile than WT or S1943A. (A, i) <t>MSCs</t> <t>transfected</t> with GFP-MIIA or GFP-MIIB on stiff matrix were photobleached along actin bundles and allowed to recover. MIIA recovered faster and thus was more mobile than MIIB. (ii) MSCs expressing GFP-MIIA on either soft 1-kPa matrix or stiff 34-kPa matrix were similarly analyzed, and MIIA in cells on soft matrix recovered more quickly. (iii) Phosphomimetic GFP-MIIA S1943D mutant recovered faster than GFP-MIIA S1943A, which was less than or equal to the behavior of GFP-MIIA on stiff matrix. Data are means ± SEM for at least five cells. (B) Fraction of insoluble MIIA is quantified by immunofluorescence of cell ghosts that were derived from Triton X-100 extraction of cells on matrices for 4 min (±SEM; n = 3). Western blotting of the insoluble fraction of the cells on matrices was also performed. Representative blot for MIIA is shown, normalized to total protein. (C) MSCs transfected with either GFP only, GFP-MIIA (WT), S1943D, or S1943A mutants were Triton X-100 extracted, and the extracted amount of MIIA (soluble) was compared with the insoluble fraction. Immunoblots show anti-MIIA blotting with β-actin as a loading control. When the amount of MIIA is normalized to β-actin, the only significant difference in the insoluble/soluble MIIA ratio is between S1943D and S1943A (P ≤ 0.05; ±SEM; n = 3). Endog., endogenous.
    Mscs, supplied by MEDIPOST, used in various techniques. Bioz Stars score: 92/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 27 article reviews
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    mscs - by Bioz Stars, 2020-08
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    93
    R&D Systems mscs
    IL-6- and <t>IL-6R-neutralizing</t> antibodies inhibit osteogenic differentiation in <t>BM-MSCs.</t> a IL-6, sIL-6R or both inhibited the extent of ARS staining in BM-MSCs. b ALP staining and activity of BM-MSCs were reduced by IL-6- and IL-6R-neutralizing antibodies. c Expression of osteoblastic marker genes in BM-MSCs was also inhibited by IL-6- and IL-6R-neutralizing antibodies. d IL-6- and IL-6R-neutralizing antibodies also inhibited STAT3 phosphorylation in BM-MSCs during osteogenic differentiation. Data are presented as the means ± SD of 15 samples per group. *Indicates P
    Mscs, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1052 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher mscs
    TRPM7 knockdown and blocking abolished the upregulation of osteogenic marker genes under shear flow. ( a ) Gene expressions of TRPM7 treated with scrambled <t>siRNA</t> (sham control) and TRPM7 siRNA were analyzed by RT-PCR. The expression of TRPM7 decreased to 20% compared to the sham control after siRNA treatment for 24 hours. ( b ) Relative gene expressions of TRPM7, Osterix, Runx2, and Dlx5 with and without intermittent shear stress. TRPM7 knockdown attenuated the upregulation of Osterix upon shear flow but with little effect on the gene expressions of Dlx5 and Runx2 compared to the sham control. ( c ) Relative gene expressions of Osterix, Dlx5, Runx2, COL1A1, ALPL, and bone morphogenetic proteins (BMP2) with and without 100 μM 2-APB additive were assayed by RT-PCR. 2-APB was able to diminish the upregulation of osteogenesis-related genes induced by three-hour IFSS. Data were normalized by the respective gene expressions of <t>MSCs</t> without FSS (static control) and presented as fold change ± SEM (n = 3). Significant difference *indicated p
    Mscs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 11487 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Corning Life Sciences mscs
    TRPM7 knockdown and blocking abolished the upregulation of osteogenic marker genes under shear flow. ( a ) Gene expressions of TRPM7 treated with scrambled <t>siRNA</t> (sham control) and TRPM7 siRNA were analyzed by RT-PCR. The expression of TRPM7 decreased to 20% compared to the sham control after siRNA treatment for 24 hours. ( b ) Relative gene expressions of TRPM7, Osterix, Runx2, and Dlx5 with and without intermittent shear stress. TRPM7 knockdown attenuated the upregulation of Osterix upon shear flow but with little effect on the gene expressions of Dlx5 and Runx2 compared to the sham control. ( c ) Relative gene expressions of Osterix, Dlx5, Runx2, COL1A1, ALPL, and bone morphogenetic proteins (BMP2) with and without 100 μM 2-APB additive were assayed by RT-PCR. 2-APB was able to diminish the upregulation of osteogenesis-related genes induced by three-hour IFSS. Data were normalized by the respective gene expressions of <t>MSCs</t> without FSS (static control) and presented as fold change ± SEM (n = 3). Significant difference *indicated p
    Mscs, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 92/100, based on 963 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 963 article reviews
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    92
    Atlanta Biologicals mscs
    TRPM7 knockdown and blocking abolished the upregulation of osteogenic marker genes under shear flow. ( a ) Gene expressions of TRPM7 treated with scrambled <t>siRNA</t> (sham control) and TRPM7 siRNA were analyzed by RT-PCR. The expression of TRPM7 decreased to 20% compared to the sham control after siRNA treatment for 24 hours. ( b ) Relative gene expressions of TRPM7, Osterix, Runx2, and Dlx5 with and without intermittent shear stress. TRPM7 knockdown attenuated the upregulation of Osterix upon shear flow but with little effect on the gene expressions of Dlx5 and Runx2 compared to the sham control. ( c ) Relative gene expressions of Osterix, Dlx5, Runx2, COL1A1, ALPL, and bone morphogenetic proteins (BMP2) with and without 100 μM 2-APB additive were assayed by RT-PCR. 2-APB was able to diminish the upregulation of osteogenesis-related genes induced by three-hour IFSS. Data were normalized by the respective gene expressions of <t>MSCs</t> without FSS (static control) and presented as fold change ± SEM (n = 3). Significant difference *indicated p
    Mscs, supplied by Atlanta Biologicals, used in various techniques. Bioz Stars score: 92/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GE Healthcare mscs
    A. Intact tendon (IT control group) displayed highly organized dense collagen fibers oriented parallel to each other with cells assembled between the fibers. B. Tendon allowed to naturally heal (NH group) had poorly organized new tissue. C. Tendon treated with fibrin gel alone (FG group) had an organized structure with round cells. D. Tendon treated with genetically engineered <t>MSCs</t> (GE group) had an organized structure of parallel bundles of extracellular matrix and spindle-shaped cells. E. CM-DiI imaging identified genetically engineered MSCs in the implantation site 3 weeks after treatment. F. Tendon that received <t>nonengineered</t> MSCs (CH3 group) had small round cells that were not highly organized, as well as an abundance of ECM.
    Mscs, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 634 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Lonza mscs human mscs
    Hierarchical clustering analysis based on the ratio of cytokine levels in acute myeloid leukemia (AML)-mesenchymal stem cell (MSC) cocultures divided by the concentration sums of AML cells and <t>MSCs</t> cultured alone; primary AML cells were derived from 51 patients and cocultured with <t>MSC24539</t> . Each horizontal row in the figure represents the observation for one patient, and the vertical columns represent the observations for the soluble mediators. Red indicates supra-additive effects in coculture. Likewise cytokines marked in red showed supra-additive effect on the total patient cohort, whereas mediators marked in blue showed higher levels in MSC cultures alone than in coculture. The patients clustered into two main groups (see right part of the figure) indicating high and low relative coculture cytokine levels, respectively. The figure also shows the distribution of various biological characteristics between the patient subsets (AML cell viability after 72 h of culture in medium alone and in coculture, proliferative capacity in medium alone and in coculture, FAB classification, genetic abnormalities, and expression of the CD34 stem cell marker).
    Mscs Human Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 91/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mscs human mscs - by Bioz Stars, 2020-08
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    92
    Cambrex mscs
    Hierarchical clustering analysis based on the ratio of cytokine levels in acute myeloid leukemia (AML)-mesenchymal stem cell (MSC) cocultures divided by the concentration sums of AML cells and <t>MSCs</t> cultured alone; primary AML cells were derived from 51 patients and cocultured with <t>MSC24539</t> . Each horizontal row in the figure represents the observation for one patient, and the vertical columns represent the observations for the soluble mediators. Red indicates supra-additive effects in coculture. Likewise cytokines marked in red showed supra-additive effect on the total patient cohort, whereas mediators marked in blue showed higher levels in MSC cultures alone than in coculture. The patients clustered into two main groups (see right part of the figure) indicating high and low relative coculture cytokine levels, respectively. The figure also shows the distribution of various biological characteristics between the patient subsets (AML cell viability after 72 h of culture in medium alone and in coculture, proliferative capacity in medium alone and in coculture, FAB classification, genetic abnormalities, and expression of the CD34 stem cell marker).
    Mscs, supplied by Cambrex, used in various techniques. Bioz Stars score: 92/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    PromoCell mscs
    Hierarchical clustering analysis based on the ratio of cytokine levels in acute myeloid leukemia (AML)-mesenchymal stem cell (MSC) cocultures divided by the concentration sums of AML cells and <t>MSCs</t> cultured alone; primary AML cells were derived from 51 patients and cocultured with <t>MSC24539</t> . Each horizontal row in the figure represents the observation for one patient, and the vertical columns represent the observations for the soluble mediators. Red indicates supra-additive effects in coculture. Likewise cytokines marked in red showed supra-additive effect on the total patient cohort, whereas mediators marked in blue showed higher levels in MSC cultures alone than in coculture. The patients clustered into two main groups (see right part of the figure) indicating high and low relative coculture cytokine levels, respectively. The figure also shows the distribution of various biological characteristics between the patient subsets (AML cell viability after 72 h of culture in medium alone and in coculture, proliferative capacity in medium alone and in coculture, FAB classification, genetic abnormalities, and expression of the CD34 stem cell marker).
    Mscs, supplied by PromoCell, used in various techniques. Bioz Stars score: 92/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 72 article reviews
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    85
    JCR Pharmaceuticals msc cm bm mscs
    Hierarchical clustering analysis based on the ratio of cytokine levels in acute myeloid leukemia (AML)-mesenchymal stem cell (MSC) cocultures divided by the concentration sums of AML cells and <t>MSCs</t> cultured alone; primary AML cells were derived from 51 patients and cocultured with <t>MSC24539</t> . Each horizontal row in the figure represents the observation for one patient, and the vertical columns represent the observations for the soluble mediators. Red indicates supra-additive effects in coculture. Likewise cytokines marked in red showed supra-additive effect on the total patient cohort, whereas mediators marked in blue showed higher levels in MSC cultures alone than in coculture. The patients clustered into two main groups (see right part of the figure) indicating high and low relative coculture cytokine levels, respectively. The figure also shows the distribution of various biological characteristics between the patient subsets (AML cell viability after 72 h of culture in medium alone and in coculture, proliferative capacity in medium alone and in coculture, FAB classification, genetic abnormalities, and expression of the CD34 stem cell marker).
    Msc Cm Bm Mscs, supplied by JCR Pharmaceuticals, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Freund-Vector mscs
    Intracellular pool of <t>prominin-1</t> is depleted upon PMA treatment A,B. HSPCs pre-cultured on <t>MSCs</t> for 2 days were incubated in the absence (control) or presence of phorbol ester PMA (PMA). After 3 (A) or 1 (B) days, cells were first cell surface-labelled for prominin-1 (red) prior to PFA-fixation and saponin-permeabilization followed by a second round of prominin-1 (green) labelling. The nuclei were stained with DAPI (blue). The labelled cells were analysed using confocal laser-scanning microscopy. A single optical x – y -plane section is shown. Scale bar, 5 µm [(A) and (B)]. C. After 3 days of incubation without (left panel) or with (right panel) PMA the number (indicated above the bar) of prominin-1-positive cells (open areas) was reduced, whereas, the MFI was slightly increased as analysed by flow cytometry. Appropriate isotype-matching control is depicted (grey filled areas). D. The ratio of intracellular (I) versus cell surface (S) fluorescence intensity of prominin-1 was quantified using CellProfiler 2.0 software. Results are the mean ± SEM. ( n = 60 cells; three independent donors). Note the reduction of the intracellular pool of prominin-1 upon 1 or 3 days (D) of PMA treatment.
    Mscs, supplied by Freund-Vector, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson mscs
    Intracellular pool of <t>prominin-1</t> is depleted upon PMA treatment A,B. HSPCs pre-cultured on <t>MSCs</t> for 2 days were incubated in the absence (control) or presence of phorbol ester PMA (PMA). After 3 (A) or 1 (B) days, cells were first cell surface-labelled for prominin-1 (red) prior to PFA-fixation and saponin-permeabilization followed by a second round of prominin-1 (green) labelling. The nuclei were stained with DAPI (blue). The labelled cells were analysed using confocal laser-scanning microscopy. A single optical x – y -plane section is shown. Scale bar, 5 µm [(A) and (B)]. C. After 3 days of incubation without (left panel) or with (right panel) PMA the number (indicated above the bar) of prominin-1-positive cells (open areas) was reduced, whereas, the MFI was slightly increased as analysed by flow cytometry. Appropriate isotype-matching control is depicted (grey filled areas). D. The ratio of intracellular (I) versus cell surface (S) fluorescence intensity of prominin-1 was quantified using CellProfiler 2.0 software. Results are the mean ± SEM. ( n = 60 cells; three independent donors). Note the reduction of the intracellular pool of prominin-1 upon 1 or 3 days (D) of PMA treatment.
    Mscs, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 3963 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Epitomics mscs
    Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC <t>MSCs)</t> to produce <t>IDO1</t> (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p
    Mscs, supplied by Epitomics, used in various techniques. Bioz Stars score: 92/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Merck KGaA mscs
    Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC <t>MSCs)</t> to produce <t>IDO1</t> (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p
    Mscs, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Osiris Therapeutics mscs
    Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC <t>MSCs)</t> to produce <t>IDO1</t> (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p
    Mscs, supplied by Osiris Therapeutics, used in various techniques. Bioz Stars score: 93/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher at mscs
    Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC <t>MSCs)</t> to produce <t>IDO1</t> (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p
    At Mscs, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 188 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Mesoblast Ltd mscs
    Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC <t>MSCs)</t> to produce <t>IDO1</t> (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p
    Mscs, supplied by Mesoblast Ltd, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    MEDIPOST hucb mscs hucb mscs
    Typical presence of <t>hUCB-MSCs</t> and expression of inflammatory factors following the transplantation of hUCB-MSCs into ICH rats. (A) Three days after the transplantation, hUCB-MSCs were identified by staining with human nuclei antibody (HuNu, red). Confocal micrograph of a brain section show the HuNu-positive cells in peri-infarct regions. The expressions of TNF-α (green) (B), COX-2 (green) (C), CD11b (green) (D), and MPO (green) (E) were detected in the perihemorrhagic regions. The number of TNF-α-positive cells (F), COX-2 positive cells (G), CD11b-reactive microglia (H), and MPO-reactive neutrophils (I) decreased in the hUCB-MSCs-treated group compared with the controls (no significant differences). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM; n=3 per group.
    Hucb Mscs Hucb Mscs, supplied by MEDIPOST, used in various techniques. Bioz Stars score: 89/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    hucb mscs hucb mscs - by Bioz Stars, 2020-08
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    85
    Lonza bone marrow mscs mscs
    Typical presence of <t>hUCB-MSCs</t> and expression of inflammatory factors following the transplantation of hUCB-MSCs into ICH rats. (A) Three days after the transplantation, hUCB-MSCs were identified by staining with human nuclei antibody (HuNu, red). Confocal micrograph of a brain section show the HuNu-positive cells in peri-infarct regions. The expressions of TNF-α (green) (B), COX-2 (green) (C), CD11b (green) (D), and MPO (green) (E) were detected in the perihemorrhagic regions. The number of TNF-α-positive cells (F), COX-2 positive cells (G), CD11b-reactive microglia (H), and MPO-reactive neutrophils (I) decreased in the hUCB-MSCs-treated group compared with the controls (no significant differences). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM; n=3 per group.
    Bone Marrow Mscs Mscs, supplied by Lonza, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bone marrow mscs mscs/product/Lonza
    Average 85 stars, based on 8 article reviews
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    bone marrow mscs mscs - by Bioz Stars, 2020-08
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    85
    Thermo Fisher rat bm msc cbm msc
    Typical presence of <t>hUCB-MSCs</t> and expression of inflammatory factors following the transplantation of hUCB-MSCs into ICH rats. (A) Three days after the transplantation, hUCB-MSCs were identified by staining with human nuclei antibody (HuNu, red). Confocal micrograph of a brain section show the HuNu-positive cells in peri-infarct regions. The expressions of TNF-α (green) (B), COX-2 (green) (C), CD11b (green) (D), and MPO (green) (E) were detected in the perihemorrhagic regions. The number of TNF-α-positive cells (F), COX-2 positive cells (G), CD11b-reactive microglia (H), and MPO-reactive neutrophils (I) decreased in the hUCB-MSCs-treated group compared with the controls (no significant differences). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM; n=3 per group.
    Rat Bm Msc Cbm Msc, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rat bm msc cbm msc/product/Thermo Fisher
    Average 85 stars, based on 1 article reviews
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    Image Search Results


    CTLA-4 affected the tumourigenic capacity of melanoma cells. C57BL/6 mice were inoculated with (A) B16-F0 cells and (B) B16-F1 cells for 7 days, separately. C57BL/6 mice were then administered with CTLA-4 inhibitor. The tumour size of the melanoma was monitored. The residual ALDH + MSCs within the tumour were analysed by flow cytometry in a (C) B16-F0 mouse model and (D) B16-F1 mouse model. **P

    Journal: Oncology Letters

    Article Title: Potential function of CTLA-4 in the tumourigenic capacity of melanoma stem cells

    doi: 10.3892/ol.2018.9354

    Figure Lengend Snippet: CTLA-4 affected the tumourigenic capacity of melanoma cells. C57BL/6 mice were inoculated with (A) B16-F0 cells and (B) B16-F1 cells for 7 days, separately. C57BL/6 mice were then administered with CTLA-4 inhibitor. The tumour size of the melanoma was monitored. The residual ALDH + MSCs within the tumour were analysed by flow cytometry in a (C) B16-F0 mouse model and (D) B16-F1 mouse model. **P

    Article Snippet: To investigate the expression of CTLA-4 in MSCs, the ALDEFLUOR kit (Stemcell Technologies, Inc., Vancouver, BC, Canada) was used.

    Techniques: Mouse Assay, Flow Cytometry, Cytometry

    Infused mesenchymal stem cells (MSCs) persist in spleen after ischemia/reperfusion injury (IRI). Intravenously delivered RFP-labeled MSCs were not detected in ischemic kidney at any time point ( a ), but persisted in spleen during the whole process, at least for 120 h after reperfusion ( b ). RFP-labeled MSCs (red) were detected by laser confocal microscopy. Nuclei were stained with DAPI (blue). Original magnification, × 600, n =3. ( c ) Intravenously delivered DiR-labeled MSCs (yellow and red) persisted in the spleen at least for 120 h after reperfusion. Signals were detected by IVIS in vivo imaging, n =3.

    Journal: Kidney International

    Article Title: Mesenchymal stem cells attenuate ischemic acute kidney injury by inducing regulatory T cells through splenocyte interactions

    doi: 10.1038/ki.2013.114

    Figure Lengend Snippet: Infused mesenchymal stem cells (MSCs) persist in spleen after ischemia/reperfusion injury (IRI). Intravenously delivered RFP-labeled MSCs were not detected in ischemic kidney at any time point ( a ), but persisted in spleen during the whole process, at least for 120 h after reperfusion ( b ). RFP-labeled MSCs (red) were detected by laser confocal microscopy. Nuclei were stained with DAPI (blue). Original magnification, × 600, n =3. ( c ) Intravenously delivered DiR-labeled MSCs (yellow and red) persisted in the spleen at least for 120 h after reperfusion. Signals were detected by IVIS in vivo imaging, n =3.

    Article Snippet: Cell preparation RFP-labeled mouse bone marrow–derived MSCs were obtained from Cyagen Biosciences (Cyagen Biosciences, Sunnyvale, CA).

    Techniques: Labeling, Confocal Microscopy, Staining, In Vivo Imaging

    MIIA is more assembled on stiff matrix than soft, and the phosphomimetic S1943D is more soluble and mobile than WT or S1943A. (A, i) MSCs transfected with GFP-MIIA or GFP-MIIB on stiff matrix were photobleached along actin bundles and allowed to recover. MIIA recovered faster and thus was more mobile than MIIB. (ii) MSCs expressing GFP-MIIA on either soft 1-kPa matrix or stiff 34-kPa matrix were similarly analyzed, and MIIA in cells on soft matrix recovered more quickly. (iii) Phosphomimetic GFP-MIIA S1943D mutant recovered faster than GFP-MIIA S1943A, which was less than or equal to the behavior of GFP-MIIA on stiff matrix. Data are means ± SEM for at least five cells. (B) Fraction of insoluble MIIA is quantified by immunofluorescence of cell ghosts that were derived from Triton X-100 extraction of cells on matrices for 4 min (±SEM; n = 3). Western blotting of the insoluble fraction of the cells on matrices was also performed. Representative blot for MIIA is shown, normalized to total protein. (C) MSCs transfected with either GFP only, GFP-MIIA (WT), S1943D, or S1943A mutants were Triton X-100 extracted, and the extracted amount of MIIA (soluble) was compared with the insoluble fraction. Immunoblots show anti-MIIA blotting with β-actin as a loading control. When the amount of MIIA is normalized to β-actin, the only significant difference in the insoluble/soluble MIIA ratio is between S1943D and S1943A (P ≤ 0.05; ±SEM; n = 3). Endog., endogenous.

    Journal: The Journal of Cell Biology

    Article Title: Crawling from soft to stiff matrix polarizes the cytoskeleton and phosphoregulates myosin-II heavy chain

    doi: 10.1083/jcb.201205056

    Figure Lengend Snippet: MIIA is more assembled on stiff matrix than soft, and the phosphomimetic S1943D is more soluble and mobile than WT or S1943A. (A, i) MSCs transfected with GFP-MIIA or GFP-MIIB on stiff matrix were photobleached along actin bundles and allowed to recover. MIIA recovered faster and thus was more mobile than MIIB. (ii) MSCs expressing GFP-MIIA on either soft 1-kPa matrix or stiff 34-kPa matrix were similarly analyzed, and MIIA in cells on soft matrix recovered more quickly. (iii) Phosphomimetic GFP-MIIA S1943D mutant recovered faster than GFP-MIIA S1943A, which was less than or equal to the behavior of GFP-MIIA on stiff matrix. Data are means ± SEM for at least five cells. (B) Fraction of insoluble MIIA is quantified by immunofluorescence of cell ghosts that were derived from Triton X-100 extraction of cells on matrices for 4 min (±SEM; n = 3). Western blotting of the insoluble fraction of the cells on matrices was also performed. Representative blot for MIIA is shown, normalized to total protein. (C) MSCs transfected with either GFP only, GFP-MIIA (WT), S1943D, or S1943A mutants were Triton X-100 extracted, and the extracted amount of MIIA (soluble) was compared with the insoluble fraction. Immunoblots show anti-MIIA blotting with β-actin as a loading control. When the amount of MIIA is normalized to β-actin, the only significant difference in the insoluble/soluble MIIA ratio is between S1943D and S1943A (P ≤ 0.05; ±SEM; n = 3). Endog., endogenous.

    Article Snippet: Constructs were transfected via electroporation following the recommended procedure of the kit for MSCs (Nucleofector; Lonza) or by using Lipofectamine LTX (Invitrogen) with plus reagent using 0.5 g DNA per well in a 6-well plate.

    Techniques: Transfection, Expressing, Mutagenesis, Immunofluorescence, Derivative Assay, Western Blot

    Durotaxis and MIIB polarization are disrupted by MIIA S1943D overexpression and are maximal for WT levels of MIIB expression and pS1943. (A) MSCs on gradient gels transfected with WT GFP-MIIA show a durotaxis index similar to nontransfected, WT cells (yellow band). The S1943A mutant shows a reduced durotaxis index, and the S1943D mutant shows no significant durotaxis above background. Data are means ± SEM for ≥12 cells. (B) The same transfected MSCs were immunostained for MIIB, which polarizes in GFP-MIIA cells on stiff matrix (34 kPa) but does not on soft matrix (1 kPa) similar to nontransfected, WT cells ( Fig. 1 ). The S1943A mutant shows a modest increase in MIIB polarization even on soft matrix, where these cells tend to spread more than any other cell ( Fig. S5 ), as is found on stiff matrix. The S1943D mutant shows no significant MIIB polarization on soft or stiff matrix, even though these cells tend to spread on stiff matrix more so than any other cell (Fig. S5). The blue band is the range of durotaxis index when there is no durotaxis per Fig. 3 C . (C) Summary of all durotaxis index and polarization data from WT, KD, and overexpression experiments shows that WT cells are optimized for durotaxis and polarization. For only the data point involving transfection with S1943D, the S1943D is considered equivalent to pS1943 and considered part of the percentage of pS1943 in the graph. The surface plot illustrates the sensitivity of durotaxis and MIIB polarization to both MIIB expression level and the percentage of MIIA that is phosphorylated at S1943 within the cell. Circles are red for the durotaxis index, whereas squares are black for rear/front fluorescence. Data are means ± SEM for ≥20 cells among three independent experiments. Bars, 50 µm.

    Journal: The Journal of Cell Biology

    Article Title: Crawling from soft to stiff matrix polarizes the cytoskeleton and phosphoregulates myosin-II heavy chain

    doi: 10.1083/jcb.201205056

    Figure Lengend Snippet: Durotaxis and MIIB polarization are disrupted by MIIA S1943D overexpression and are maximal for WT levels of MIIB expression and pS1943. (A) MSCs on gradient gels transfected with WT GFP-MIIA show a durotaxis index similar to nontransfected, WT cells (yellow band). The S1943A mutant shows a reduced durotaxis index, and the S1943D mutant shows no significant durotaxis above background. Data are means ± SEM for ≥12 cells. (B) The same transfected MSCs were immunostained for MIIB, which polarizes in GFP-MIIA cells on stiff matrix (34 kPa) but does not on soft matrix (1 kPa) similar to nontransfected, WT cells ( Fig. 1 ). The S1943A mutant shows a modest increase in MIIB polarization even on soft matrix, where these cells tend to spread more than any other cell ( Fig. S5 ), as is found on stiff matrix. The S1943D mutant shows no significant MIIB polarization on soft or stiff matrix, even though these cells tend to spread on stiff matrix more so than any other cell (Fig. S5). The blue band is the range of durotaxis index when there is no durotaxis per Fig. 3 C . (C) Summary of all durotaxis index and polarization data from WT, KD, and overexpression experiments shows that WT cells are optimized for durotaxis and polarization. For only the data point involving transfection with S1943D, the S1943D is considered equivalent to pS1943 and considered part of the percentage of pS1943 in the graph. The surface plot illustrates the sensitivity of durotaxis and MIIB polarization to both MIIB expression level and the percentage of MIIA that is phosphorylated at S1943 within the cell. Circles are red for the durotaxis index, whereas squares are black for rear/front fluorescence. Data are means ± SEM for ≥20 cells among three independent experiments. Bars, 50 µm.

    Article Snippet: Constructs were transfected via electroporation following the recommended procedure of the kit for MSCs (Nucleofector; Lonza) or by using Lipofectamine LTX (Invitrogen) with plus reagent using 0.5 g DNA per well in a 6-well plate.

    Techniques: Over Expression, Expressing, Transfection, Mutagenesis, Fluorescence

    IL-6- and IL-6R-neutralizing antibodies inhibit osteogenic differentiation in BM-MSCs. a IL-6, sIL-6R or both inhibited the extent of ARS staining in BM-MSCs. b ALP staining and activity of BM-MSCs were reduced by IL-6- and IL-6R-neutralizing antibodies. c Expression of osteoblastic marker genes in BM-MSCs was also inhibited by IL-6- and IL-6R-neutralizing antibodies. d IL-6- and IL-6R-neutralizing antibodies also inhibited STAT3 phosphorylation in BM-MSCs during osteogenic differentiation. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Journal: Stem Cell Research & Therapy

    Article Title: Interleukin-6/interleukin-6 receptor complex promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells

    doi: 10.1186/s13287-017-0766-0

    Figure Lengend Snippet: IL-6- and IL-6R-neutralizing antibodies inhibit osteogenic differentiation in BM-MSCs. a IL-6, sIL-6R or both inhibited the extent of ARS staining in BM-MSCs. b ALP staining and activity of BM-MSCs were reduced by IL-6- and IL-6R-neutralizing antibodies. c Expression of osteoblastic marker genes in BM-MSCs was also inhibited by IL-6- and IL-6R-neutralizing antibodies. d IL-6- and IL-6R-neutralizing antibodies also inhibited STAT3 phosphorylation in BM-MSCs during osteogenic differentiation. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Article Snippet: Recombinant human IL-6 protein (100 ng/ml), recombinant human sIL-6R protein (100 ng/ml), anti-IL-6 antibody (5 μg/ml) or anti-IL-6R antibody (5 μg/ml) was added to determine the roles of IL-6/IL-6R in the osteogenic differentiation of MSCs (all from R & D Systems).

    Techniques: Staining, ALP Assay, Activity Assay, Expressing, Marker

    IL-6 and IL-6R stimulate each other’s expression in BM-MSCs. a IL-6 secretion in BM-MSCs was increased by exogenous sIL-6R but decreased by IL-6R-neutralizing antibody during osteogenic differentiation. b Western blotting results showed that mIL-6R expression in BM-MSCs was induced by exogenous IL-6 but reduced by IL-6-neutralizing antibody. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Journal: Stem Cell Research & Therapy

    Article Title: Interleukin-6/interleukin-6 receptor complex promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells

    doi: 10.1186/s13287-017-0766-0

    Figure Lengend Snippet: IL-6 and IL-6R stimulate each other’s expression in BM-MSCs. a IL-6 secretion in BM-MSCs was increased by exogenous sIL-6R but decreased by IL-6R-neutralizing antibody during osteogenic differentiation. b Western blotting results showed that mIL-6R expression in BM-MSCs was induced by exogenous IL-6 but reduced by IL-6-neutralizing antibody. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Article Snippet: Recombinant human IL-6 protein (100 ng/ml), recombinant human sIL-6R protein (100 ng/ml), anti-IL-6 antibody (5 μg/ml) or anti-IL-6R antibody (5 μg/ml) was added to determine the roles of IL-6/IL-6R in the osteogenic differentiation of MSCs (all from R & D Systems).

    Techniques: Expressing, Western Blot

    IL-6 and IL-6R expression in BM-MSCs during osteogenic differentiation. a Expression of IL-6 and IL-6R genes increased during osteogenic differentiation and peaked on day 10 or 14 of induction. b ELISA results showing that IL-6 secretion increases during osteogenic differentiation in BM-MSCs. Western blotting results showing that IL-6R expression in BM-MSCs peaked on day 14 of induction. c Results of flow cytometry showing that mIL-6R expression increases during osteogenic differentiation in BM-MSCs. d IL-6 and IL-6R expression are positively correlated with ARS staining results in BM-MSCs. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Journal: Stem Cell Research & Therapy

    Article Title: Interleukin-6/interleukin-6 receptor complex promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells

    doi: 10.1186/s13287-017-0766-0

    Figure Lengend Snippet: IL-6 and IL-6R expression in BM-MSCs during osteogenic differentiation. a Expression of IL-6 and IL-6R genes increased during osteogenic differentiation and peaked on day 10 or 14 of induction. b ELISA results showing that IL-6 secretion increases during osteogenic differentiation in BM-MSCs. Western blotting results showing that IL-6R expression in BM-MSCs peaked on day 14 of induction. c Results of flow cytometry showing that mIL-6R expression increases during osteogenic differentiation in BM-MSCs. d IL-6 and IL-6R expression are positively correlated with ARS staining results in BM-MSCs. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Article Snippet: Recombinant human IL-6 protein (100 ng/ml), recombinant human sIL-6R protein (100 ng/ml), anti-IL-6 antibody (5 μg/ml) or anti-IL-6R antibody (5 μg/ml) was added to determine the roles of IL-6/IL-6R in the osteogenic differentiation of MSCs (all from R & D Systems).

    Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Flow Cytometry, Cytometry, Staining

    Exogenous IL-6 and sIL-6R promote osteogenic differentiation in BM-MSCs. a Extent of ARS staining in BM-MSCs was clearly increased by exogenous IL-6, sIL-6R or both. b ALP staining and activity of BM-MSCs were noticeably increased by exogenous IL-6 and sIL-6R. c Expression of osteoblastic marker genes in BM-MSCs was also promoted by exogenous IL-6 and sIL-6R. d STAT3 phosphorylation was markedly increased in response to stimulation with exogenous IL-6 and sIL-6R. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Journal: Stem Cell Research & Therapy

    Article Title: Interleukin-6/interleukin-6 receptor complex promotes osteogenic differentiation of bone marrow-derived mesenchymal stem cells

    doi: 10.1186/s13287-017-0766-0

    Figure Lengend Snippet: Exogenous IL-6 and sIL-6R promote osteogenic differentiation in BM-MSCs. a Extent of ARS staining in BM-MSCs was clearly increased by exogenous IL-6, sIL-6R or both. b ALP staining and activity of BM-MSCs were noticeably increased by exogenous IL-6 and sIL-6R. c Expression of osteoblastic marker genes in BM-MSCs was also promoted by exogenous IL-6 and sIL-6R. d STAT3 phosphorylation was markedly increased in response to stimulation with exogenous IL-6 and sIL-6R. Data are presented as the means ± SD of 15 samples per group. *Indicates P

    Article Snippet: Recombinant human IL-6 protein (100 ng/ml), recombinant human sIL-6R protein (100 ng/ml), anti-IL-6 antibody (5 μg/ml) or anti-IL-6R antibody (5 μg/ml) was added to determine the roles of IL-6/IL-6R in the osteogenic differentiation of MSCs (all from R & D Systems).

    Techniques: Staining, ALP Assay, Activity Assay, Expressing, Marker

    TRPM7 knockdown and blocking abolished the upregulation of osteogenic marker genes under shear flow. ( a ) Gene expressions of TRPM7 treated with scrambled siRNA (sham control) and TRPM7 siRNA were analyzed by RT-PCR. The expression of TRPM7 decreased to 20% compared to the sham control after siRNA treatment for 24 hours. ( b ) Relative gene expressions of TRPM7, Osterix, Runx2, and Dlx5 with and without intermittent shear stress. TRPM7 knockdown attenuated the upregulation of Osterix upon shear flow but with little effect on the gene expressions of Dlx5 and Runx2 compared to the sham control. ( c ) Relative gene expressions of Osterix, Dlx5, Runx2, COL1A1, ALPL, and bone morphogenetic proteins (BMP2) with and without 100 μM 2-APB additive were assayed by RT-PCR. 2-APB was able to diminish the upregulation of osteogenesis-related genes induced by three-hour IFSS. Data were normalized by the respective gene expressions of MSCs without FSS (static control) and presented as fold change ± SEM (n = 3). Significant difference *indicated p

    Journal: Scientific Reports

    Article Title: Mechanosensitive TRPM7 mediates shear stress and modulates osteogenic differentiation of mesenchymal stromal cells through Osterix pathway

    doi: 10.1038/srep16522

    Figure Lengend Snippet: TRPM7 knockdown and blocking abolished the upregulation of osteogenic marker genes under shear flow. ( a ) Gene expressions of TRPM7 treated with scrambled siRNA (sham control) and TRPM7 siRNA were analyzed by RT-PCR. The expression of TRPM7 decreased to 20% compared to the sham control after siRNA treatment for 24 hours. ( b ) Relative gene expressions of TRPM7, Osterix, Runx2, and Dlx5 with and without intermittent shear stress. TRPM7 knockdown attenuated the upregulation of Osterix upon shear flow but with little effect on the gene expressions of Dlx5 and Runx2 compared to the sham control. ( c ) Relative gene expressions of Osterix, Dlx5, Runx2, COL1A1, ALPL, and bone morphogenetic proteins (BMP2) with and without 100 μM 2-APB additive were assayed by RT-PCR. 2-APB was able to diminish the upregulation of osteogenesis-related genes induced by three-hour IFSS. Data were normalized by the respective gene expressions of MSCs without FSS (static control) and presented as fold change ± SEM (n = 3). Significant difference *indicated p

    Article Snippet: siRNA transfection of TRPM7 siRNA with sequences targeting TRPM7 was transfected into MSCs according to the manufacturer’s instructions (Invitrogen).

    Techniques: Blocking Assay, Marker, Flow Cytometry, Reverse Transcription Polymerase Chain Reaction, Expressing

    TRPM7 modulated cytoskeletal arrangement of MSCs in the early stage of osteogenic differentiation. ( a ) Relative gene expressions of MSCs on the third day of osteogenic differentiation in static condition with or without 20 μM 2-APB. Data were normalized by the respective gene expressions of MSCs without 2-APB (control). 2-APB blocking of TRPM7 downregulated gene expressions of Osterix, Dlx5, COL1A1, and ALPL. The blocking had no effect on gene expressions Runx2 and OCN. ( b ) Immunofluorescent staining of osteoblast-engaged MSCs on the third day after osteogenic induction in static condition with (middle) and without 20 μM 2-APB (left, as control) added in induction medium, as well as with 2-APB added in induction medium and cells were challenged by 10 nM bradykinin (BK) intermittently for 115 minutes (right). Scale bar = 100 μm. ( c ) Mean fluorescence intensities of vinculin and F-actin of the three groups (N = 34 for each group). Expressions of vinculin and F-actin increased and exhibited more scattered and less localized distribution in the TRPM7-inhibited group. Challenge of 10 nM Bradykinin intermittently rescued the effects caused by 2-APB. Data were normalized by control and presented as mean ± SEM. Significant difference *indicated p

    Journal: Scientific Reports

    Article Title: Mechanosensitive TRPM7 mediates shear stress and modulates osteogenic differentiation of mesenchymal stromal cells through Osterix pathway

    doi: 10.1038/srep16522

    Figure Lengend Snippet: TRPM7 modulated cytoskeletal arrangement of MSCs in the early stage of osteogenic differentiation. ( a ) Relative gene expressions of MSCs on the third day of osteogenic differentiation in static condition with or without 20 μM 2-APB. Data were normalized by the respective gene expressions of MSCs without 2-APB (control). 2-APB blocking of TRPM7 downregulated gene expressions of Osterix, Dlx5, COL1A1, and ALPL. The blocking had no effect on gene expressions Runx2 and OCN. ( b ) Immunofluorescent staining of osteoblast-engaged MSCs on the third day after osteogenic induction in static condition with (middle) and without 20 μM 2-APB (left, as control) added in induction medium, as well as with 2-APB added in induction medium and cells were challenged by 10 nM bradykinin (BK) intermittently for 115 minutes (right). Scale bar = 100 μm. ( c ) Mean fluorescence intensities of vinculin and F-actin of the three groups (N = 34 for each group). Expressions of vinculin and F-actin increased and exhibited more scattered and less localized distribution in the TRPM7-inhibited group. Challenge of 10 nM Bradykinin intermittently rescued the effects caused by 2-APB. Data were normalized by control and presented as mean ± SEM. Significant difference *indicated p

    Article Snippet: siRNA transfection of TRPM7 siRNA with sequences targeting TRPM7 was transfected into MSCs according to the manufacturer’s instructions (Invitrogen).

    Techniques: Blocking Assay, Staining, Fluorescence

    A. Intact tendon (IT control group) displayed highly organized dense collagen fibers oriented parallel to each other with cells assembled between the fibers. B. Tendon allowed to naturally heal (NH group) had poorly organized new tissue. C. Tendon treated with fibrin gel alone (FG group) had an organized structure with round cells. D. Tendon treated with genetically engineered MSCs (GE group) had an organized structure of parallel bundles of extracellular matrix and spindle-shaped cells. E. CM-DiI imaging identified genetically engineered MSCs in the implantation site 3 weeks after treatment. F. Tendon that received nonengineered MSCs (CH3 group) had small round cells that were not highly organized, as well as an abundance of ECM.

    Journal: Journal of orthopaedic research : official publication of the Orthopaedic Research Society

    Article Title: Smad8/BMP2-Engineered Mesenchymal Stem Cells Induce Accelerated Recovery of the Biomechanical Properties of the Achilles Tendon

    doi: 10.1002/jor.22167

    Figure Lengend Snippet: A. Intact tendon (IT control group) displayed highly organized dense collagen fibers oriented parallel to each other with cells assembled between the fibers. B. Tendon allowed to naturally heal (NH group) had poorly organized new tissue. C. Tendon treated with fibrin gel alone (FG group) had an organized structure with round cells. D. Tendon treated with genetically engineered MSCs (GE group) had an organized structure of parallel bundles of extracellular matrix and spindle-shaped cells. E. CM-DiI imaging identified genetically engineered MSCs in the implantation site 3 weeks after treatment. F. Tendon that received nonengineered MSCs (CH3 group) had small round cells that were not highly organized, as well as an abundance of ECM.

    Article Snippet: The injured tendons were then sutured together or given implants of genetically engineered MSCs (GE group), nonengineered MSCs (CH3 group), or fibrin gel containing no cells (FG group).

    Techniques: Imaging

    Hierarchical clustering analysis based on the ratio of cytokine levels in acute myeloid leukemia (AML)-mesenchymal stem cell (MSC) cocultures divided by the concentration sums of AML cells and MSCs cultured alone; primary AML cells were derived from 51 patients and cocultured with MSC24539 . Each horizontal row in the figure represents the observation for one patient, and the vertical columns represent the observations for the soluble mediators. Red indicates supra-additive effects in coculture. Likewise cytokines marked in red showed supra-additive effect on the total patient cohort, whereas mediators marked in blue showed higher levels in MSC cultures alone than in coculture. The patients clustered into two main groups (see right part of the figure) indicating high and low relative coculture cytokine levels, respectively. The figure also shows the distribution of various biological characteristics between the patient subsets (AML cell viability after 72 h of culture in medium alone and in coculture, proliferative capacity in medium alone and in coculture, FAB classification, genetic abnormalities, and expression of the CD34 stem cell marker).

    Journal: Frontiers in Immunology

    Article Title: Mesenchymal Stem Cells Support Survival and Proliferation of Primary Human Acute Myeloid Leukemia Cells through Heterogeneous Molecular Mechanisms

    doi: 10.3389/fimmu.2017.00106

    Figure Lengend Snippet: Hierarchical clustering analysis based on the ratio of cytokine levels in acute myeloid leukemia (AML)-mesenchymal stem cell (MSC) cocultures divided by the concentration sums of AML cells and MSCs cultured alone; primary AML cells were derived from 51 patients and cocultured with MSC24539 . Each horizontal row in the figure represents the observation for one patient, and the vertical columns represent the observations for the soluble mediators. Red indicates supra-additive effects in coculture. Likewise cytokines marked in red showed supra-additive effect on the total patient cohort, whereas mediators marked in blue showed higher levels in MSC cultures alone than in coculture. The patients clustered into two main groups (see right part of the figure) indicating high and low relative coculture cytokine levels, respectively. The figure also shows the distribution of various biological characteristics between the patient subsets (AML cell viability after 72 h of culture in medium alone and in coculture, proliferative capacity in medium alone and in coculture, FAB classification, genetic abnormalities, and expression of the CD34 stem cell marker).

    Article Snippet: In Vitro Expansion of MSCs Human MSCs from three healthy donors (MSC24429, MSC24539, and MSC25200) were purchased from Lonza (Cambrex BioScience, Walkersville, MD, USA).

    Techniques: Concentration Assay, Cell Culture, Derivative Assay, Expressing, Marker

    The effects of mesenchymal stem cells (MSCs) on acute myeloid leukemia (AML) cell proliferation/viability in suspension cultures and AML cells tested in a clonogenic assay . The effect of MSCs on the in vitro proliferation [ (A) , left] and viability [ (B) , middle] of primary human AML cells derived from 51 consecutive patients was examined. The AML cells were then cocultured with normal MSC24539. The MSCs were initially cultured for 3 days to allow them to establish adherent in vitro proliferation; primary human AML cells were thereafter added and (A) leukemia cell proliferation was assayed as 3 H-thymidine incorporation after additional 3 days of culture; additionally, (B) AML cell viability was assayed by the Annexin V-PI flow cytometric assay. The p -values for the statistical comparison of the overall results are given at the top of the figure. Each line represents the results for one patient. [ (C) , right] The effects of MSCs on clonogenic AML cells were also investigated. The leukemic cells were cultured either in medium alone or in transwell cocultures together with MSC24593 for 3 weeks; the number of clonogenic cells were thereafter compared for AML cells precultured in medium alone and together with MSCs. The total culture period was thus 5 weeks. The MSCs significantly increased the number of colony-forming units ( p -values are given on top of the figure) in growth media both with and without erythropoietin. The results are presented as mean of duplicate samples (average deviation from mean corresponding to 1.3 colonies and 8.7% of the total colony number). Each line represents the results for one patient.

    Journal: Frontiers in Immunology

    Article Title: Mesenchymal Stem Cells Support Survival and Proliferation of Primary Human Acute Myeloid Leukemia Cells through Heterogeneous Molecular Mechanisms

    doi: 10.3389/fimmu.2017.00106

    Figure Lengend Snippet: The effects of mesenchymal stem cells (MSCs) on acute myeloid leukemia (AML) cell proliferation/viability in suspension cultures and AML cells tested in a clonogenic assay . The effect of MSCs on the in vitro proliferation [ (A) , left] and viability [ (B) , middle] of primary human AML cells derived from 51 consecutive patients was examined. The AML cells were then cocultured with normal MSC24539. The MSCs were initially cultured for 3 days to allow them to establish adherent in vitro proliferation; primary human AML cells were thereafter added and (A) leukemia cell proliferation was assayed as 3 H-thymidine incorporation after additional 3 days of culture; additionally, (B) AML cell viability was assayed by the Annexin V-PI flow cytometric assay. The p -values for the statistical comparison of the overall results are given at the top of the figure. Each line represents the results for one patient. [ (C) , right] The effects of MSCs on clonogenic AML cells were also investigated. The leukemic cells were cultured either in medium alone or in transwell cocultures together with MSC24593 for 3 weeks; the number of clonogenic cells were thereafter compared for AML cells precultured in medium alone and together with MSCs. The total culture period was thus 5 weeks. The MSCs significantly increased the number of colony-forming units ( p -values are given on top of the figure) in growth media both with and without erythropoietin. The results are presented as mean of duplicate samples (average deviation from mean corresponding to 1.3 colonies and 8.7% of the total colony number). Each line represents the results for one patient.

    Article Snippet: In Vitro Expansion of MSCs Human MSCs from three healthy donors (MSC24429, MSC24539, and MSC25200) were purchased from Lonza (Cambrex BioScience, Walkersville, MD, USA).

    Techniques: Clonogenic Assay, In Vitro, Derivative Assay, Cell Culture, Flow Cytometry

    Effects of mesenchymal stem cells (MSCs) on primary human acute myeloid leukemia (AML) cells treated with cytarabine; (A) effect of 50 nM cytarabine-exposure on AML cell viability in transwell cocultures and (B) effect of 500 nM cytarabine tested in direct-contact cocultures . Primary AML cells derived from the same 10 patients were included in all these studies, and MSC24539 was used in all experiments. The AML cell viability was analyzed by flow cytometry. [ (A) , transwell cocultures] For each patient, we compared cultures containing AML cells alone (AML) or AML cells plus MSCs (AML + MSC). The three figures from left to the middle right show (i) AML cell viability for cells cultured in medium alone with or without MSCs; (ii) AML cell viability in the presence of 50 nM cytarabine for leukemic cells cultured with and without MSCs; and (iii) a comparison of the medium culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; medium alone) and MSC culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; MSCs being added to both cultures). The results for the five patients (solid lines) for which 50 nM cytarabine exhibited a proapoptotic effect in the presence of MSCs are indicated. The results for one responding patient are presented in detail (right part of the figure; i/ii: drug-free control, iii/iv: 50 nM cytarabine without and with MSCs, respectively); the percentage of viable cells (population at the lower left) is indicated in the figures for each of the four cultures. [ (B) , direct-contact cultures] The left part of the figure compares the medium culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; medium alone) and MSC culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; MSCs being added to both cultures) when testing 500 nM cytarabine. The right part of the figure shows the percentage of viable primary AML cells (10 patients tested, one patient not tested for AML cells + cytarabine + MSC) cultured in medium alone, together with MSCs, in medium containing 500 nM cytarabine, and together with 500 nM cytarabine and MSCs.

    Journal: Frontiers in Immunology

    Article Title: Mesenchymal Stem Cells Support Survival and Proliferation of Primary Human Acute Myeloid Leukemia Cells through Heterogeneous Molecular Mechanisms

    doi: 10.3389/fimmu.2017.00106

    Figure Lengend Snippet: Effects of mesenchymal stem cells (MSCs) on primary human acute myeloid leukemia (AML) cells treated with cytarabine; (A) effect of 50 nM cytarabine-exposure on AML cell viability in transwell cocultures and (B) effect of 500 nM cytarabine tested in direct-contact cocultures . Primary AML cells derived from the same 10 patients were included in all these studies, and MSC24539 was used in all experiments. The AML cell viability was analyzed by flow cytometry. [ (A) , transwell cocultures] For each patient, we compared cultures containing AML cells alone (AML) or AML cells plus MSCs (AML + MSC). The three figures from left to the middle right show (i) AML cell viability for cells cultured in medium alone with or without MSCs; (ii) AML cell viability in the presence of 50 nM cytarabine for leukemic cells cultured with and without MSCs; and (iii) a comparison of the medium culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; medium alone) and MSC culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; MSCs being added to both cultures). The results for the five patients (solid lines) for which 50 nM cytarabine exhibited a proapoptotic effect in the presence of MSCs are indicated. The results for one responding patient are presented in detail (right part of the figure; i/ii: drug-free control, iii/iv: 50 nM cytarabine without and with MSCs, respectively); the percentage of viable cells (population at the lower left) is indicated in the figures for each of the four cultures. [ (B) , direct-contact cultures] The left part of the figure compares the medium culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; medium alone) and MSC culture ratio (i.e., viable cells in cytarabine-containing cultures versus drug-free controls; MSCs being added to both cultures) when testing 500 nM cytarabine. The right part of the figure shows the percentage of viable primary AML cells (10 patients tested, one patient not tested for AML cells + cytarabine + MSC) cultured in medium alone, together with MSCs, in medium containing 500 nM cytarabine, and together with 500 nM cytarabine and MSCs.

    Article Snippet: In Vitro Expansion of MSCs Human MSCs from three healthy donors (MSC24429, MSC24539, and MSC25200) were purchased from Lonza (Cambrex BioScience, Walkersville, MD, USA).

    Techniques: Derivative Assay, Flow Cytometry, Cytometry, Cell Culture

    The effect of normal bone marrow mesenchymal stem cells (MSCs) on in vitro proliferation and viability of primary human acute myeloid leukemia (AML) cells; a comparison of MSCs derived from three healthy donors (MSC24429, MSC24539, MSC25200) . Primary human AML cells derived from 18 consecutive/unselected leukemia patients were cocultured with normal MSCs from the three donors. The MSCs were initially cultured for 3 days to allow them to establish adherent in vitro proliferation; primary human AML cells were then added and both leukemia cell proliferation and viability were assayed after additional 3 days of culture. [ (A) , upper part] Proliferation was assayed as 3 H-thymidine incorporation. The results for each patient are presented as nuclear thymidine incorporation (counts per minute, cpm). The p -value for the statistical comparison of the overall results is given at the top of the figure for each of the individual MSC. Each line represents the results for one patient. [ (B) , lower part] Leukemia cell viability was assayed by the Annexin V-PI flow cytometric assay. The p -value for the statistical comparison of the overall results for each of the MSCs is given at the top of the figure for the MSCs. Each line represents the results for one patient.

    Journal: Frontiers in Immunology

    Article Title: Mesenchymal Stem Cells Support Survival and Proliferation of Primary Human Acute Myeloid Leukemia Cells through Heterogeneous Molecular Mechanisms

    doi: 10.3389/fimmu.2017.00106

    Figure Lengend Snippet: The effect of normal bone marrow mesenchymal stem cells (MSCs) on in vitro proliferation and viability of primary human acute myeloid leukemia (AML) cells; a comparison of MSCs derived from three healthy donors (MSC24429, MSC24539, MSC25200) . Primary human AML cells derived from 18 consecutive/unselected leukemia patients were cocultured with normal MSCs from the three donors. The MSCs were initially cultured for 3 days to allow them to establish adherent in vitro proliferation; primary human AML cells were then added and both leukemia cell proliferation and viability were assayed after additional 3 days of culture. [ (A) , upper part] Proliferation was assayed as 3 H-thymidine incorporation. The results for each patient are presented as nuclear thymidine incorporation (counts per minute, cpm). The p -value for the statistical comparison of the overall results is given at the top of the figure for each of the individual MSC. Each line represents the results for one patient. [ (B) , lower part] Leukemia cell viability was assayed by the Annexin V-PI flow cytometric assay. The p -value for the statistical comparison of the overall results for each of the MSCs is given at the top of the figure for the MSCs. Each line represents the results for one patient.

    Article Snippet: In Vitro Expansion of MSCs Human MSCs from three healthy donors (MSC24429, MSC24539, and MSC25200) were purchased from Lonza (Cambrex BioScience, Walkersville, MD, USA).

    Techniques: In Vitro, Derivative Assay, Cell Culture, Flow Cytometry

    Intracellular pool of prominin-1 is depleted upon PMA treatment A,B. HSPCs pre-cultured on MSCs for 2 days were incubated in the absence (control) or presence of phorbol ester PMA (PMA). After 3 (A) or 1 (B) days, cells were first cell surface-labelled for prominin-1 (red) prior to PFA-fixation and saponin-permeabilization followed by a second round of prominin-1 (green) labelling. The nuclei were stained with DAPI (blue). The labelled cells were analysed using confocal laser-scanning microscopy. A single optical x – y -plane section is shown. Scale bar, 5 µm [(A) and (B)]. C. After 3 days of incubation without (left panel) or with (right panel) PMA the number (indicated above the bar) of prominin-1-positive cells (open areas) was reduced, whereas, the MFI was slightly increased as analysed by flow cytometry. Appropriate isotype-matching control is depicted (grey filled areas). D. The ratio of intracellular (I) versus cell surface (S) fluorescence intensity of prominin-1 was quantified using CellProfiler 2.0 software. Results are the mean ± SEM. ( n = 60 cells; three independent donors). Note the reduction of the intracellular pool of prominin-1 upon 1 or 3 days (D) of PMA treatment.

    Journal: EMBO Molecular Medicine

    Article Title: Haematopoietic stem cell differentiation promotes the release of prominin-1/CD133-containing membrane vesicles--a role of the endocytic-exocytic pathway

    doi: 10.1002/emmm.201100147

    Figure Lengend Snippet: Intracellular pool of prominin-1 is depleted upon PMA treatment A,B. HSPCs pre-cultured on MSCs for 2 days were incubated in the absence (control) or presence of phorbol ester PMA (PMA). After 3 (A) or 1 (B) days, cells were first cell surface-labelled for prominin-1 (red) prior to PFA-fixation and saponin-permeabilization followed by a second round of prominin-1 (green) labelling. The nuclei were stained with DAPI (blue). The labelled cells were analysed using confocal laser-scanning microscopy. A single optical x – y -plane section is shown. Scale bar, 5 µm [(A) and (B)]. C. After 3 days of incubation without (left panel) or with (right panel) PMA the number (indicated above the bar) of prominin-1-positive cells (open areas) was reduced, whereas, the MFI was slightly increased as analysed by flow cytometry. Appropriate isotype-matching control is depicted (grey filled areas). D. The ratio of intracellular (I) versus cell surface (S) fluorescence intensity of prominin-1 was quantified using CellProfiler 2.0 software. Results are the mean ± SEM. ( n = 60 cells; three independent donors). Note the reduction of the intracellular pool of prominin-1 upon 1 or 3 days (D) of PMA treatment.

    Article Snippet: Again, additional investigation is urged to reveal whether prominin-1-CMV modify the biochemistry of the recipient MSCs (Freund et al, ) as it was elegantly proposed upon the direct contact of HSPCs with osteoblasts (Gillette et al, ).

    Techniques: Cell Culture, Incubation, Staining, Confocal Laser Scanning Microscopy, Flow Cytometry, Cytometry, Fluorescence, Software

    Association of prominin-1, but not CD34, with lipid raft-membrane vesicles released by HSPCs A. One-week-old co-culture HSPC/MSC conditioned medium was subjected to differential centrifugation for 5 min at 300 × g , 20 min at 1200 × g , 30 min at 10,000 × g , 60 min at 200,000 × g and 60 min at 400,000 × g . The resulting pellets were analysed by immunoblotting for either prominin-1 (top panel, arrow) or CD34 (bottom panel, arrowhead). Proteins in the 400,000 × g supernatant (Sup.) were analysed in parallel. B. The 200,000 × g pellet recovered after differential centrifugation (panel A) was subjected to equilibrium sucrose gradient (0.1–1.2 M) centrifugation. Equal volumes of the recovered fractions and the pellet were analysed by immunoblotting for prominin-1 (arrow). C,D. Negative EM analysis of the 200,000 × g pellet revealed the presence of small membrane vesicles (∼40–80 nm; arrows) and larger dense structures (∼200–600 nm; arrowhead). E. Negative staining EM of prominin-1 immunogold-labelled membrane vesicles recovered in the 200,000 × g pellet. Scale bars, 100 nm [(C–E)]. F. Haematopoietic cells (Cells) growing for 1 week in the co-culture system and 200,000 × g pellets (200,000 × g pellet) recovered upon differential centrifugation (panel A) were lysed for 30 min at 4°C in either 0.5% Triton X-100 or Lubrol WX or without detergent (PBS) and centrifuged for 60 min at 100,000 × g . The resulting supernatants (S) and pellets (P) were analysed by immunoblotting for prominin-1 (arrows). G. Lipid composition analysis of prominin-1-CMV was performed by C 14 -acetate labelling of HSPCs co-cultured with MSCs followed by TLC analysis of either haematopoietic cells (Cells) or the released prominin-1-positive vesicles recovered by a paramagnetic isolation with mAb CD133 (prominin-1-CMV). Percentages of cholesterol (Ch) and sphingomyelin (SM) of the total lipid composition are indicated. •, sample loading point.

    Journal: EMBO Molecular Medicine

    Article Title: Haematopoietic stem cell differentiation promotes the release of prominin-1/CD133-containing membrane vesicles--a role of the endocytic-exocytic pathway

    doi: 10.1002/emmm.201100147

    Figure Lengend Snippet: Association of prominin-1, but not CD34, with lipid raft-membrane vesicles released by HSPCs A. One-week-old co-culture HSPC/MSC conditioned medium was subjected to differential centrifugation for 5 min at 300 × g , 20 min at 1200 × g , 30 min at 10,000 × g , 60 min at 200,000 × g and 60 min at 400,000 × g . The resulting pellets were analysed by immunoblotting for either prominin-1 (top panel, arrow) or CD34 (bottom panel, arrowhead). Proteins in the 400,000 × g supernatant (Sup.) were analysed in parallel. B. The 200,000 × g pellet recovered after differential centrifugation (panel A) was subjected to equilibrium sucrose gradient (0.1–1.2 M) centrifugation. Equal volumes of the recovered fractions and the pellet were analysed by immunoblotting for prominin-1 (arrow). C,D. Negative EM analysis of the 200,000 × g pellet revealed the presence of small membrane vesicles (∼40–80 nm; arrows) and larger dense structures (∼200–600 nm; arrowhead). E. Negative staining EM of prominin-1 immunogold-labelled membrane vesicles recovered in the 200,000 × g pellet. Scale bars, 100 nm [(C–E)]. F. Haematopoietic cells (Cells) growing for 1 week in the co-culture system and 200,000 × g pellets (200,000 × g pellet) recovered upon differential centrifugation (panel A) were lysed for 30 min at 4°C in either 0.5% Triton X-100 or Lubrol WX or without detergent (PBS) and centrifuged for 60 min at 100,000 × g . The resulting supernatants (S) and pellets (P) were analysed by immunoblotting for prominin-1 (arrows). G. Lipid composition analysis of prominin-1-CMV was performed by C 14 -acetate labelling of HSPCs co-cultured with MSCs followed by TLC analysis of either haematopoietic cells (Cells) or the released prominin-1-positive vesicles recovered by a paramagnetic isolation with mAb CD133 (prominin-1-CMV). Percentages of cholesterol (Ch) and sphingomyelin (SM) of the total lipid composition are indicated. •, sample loading point.

    Article Snippet: Again, additional investigation is urged to reveal whether prominin-1-CMV modify the biochemistry of the recipient MSCs (Freund et al, ) as it was elegantly proposed upon the direct contact of HSPCs with osteoblasts (Gillette et al, ).

    Techniques: Co-Culture Assay, Centrifugation, Negative Staining, Cell Culture, Thin Layer Chromatography, Isolation

    Provoked differentiation stimulated prominin-1-containing membrane vesicle release A,B. HSPCs pre-cultured either on MSCs or fibronectin-coated dishes for 2 days were incubated in the absence (A) or presence (B) of PMA. After 3 days, conditioned media were subjected to differential centrifugation as in Fig 1A . The resulting pellets were analysed by immunoblotting for prominin-1, actin and α-tubulin. Note that only 1/3 of materials were loaded for the 300 × g fraction. The amount of prominin-1 immunoreactive materials was quantified [ n = 3 (MSCs) and 4 (fibronectin)]. Results are the mean ± SEM. Ratio of prominin-1-immunoreactivity found in the 200,000 × g pellet (vesicles) versus 300 × g (cells) is indicated in bracket. Note that the provoked HSPC differentiation triggered by PMA resulted in an increase of prominin-1 in 200,000 × g pellets, which are devoid of cytoskeletal proteins under both culture conditions.

    Journal: EMBO Molecular Medicine

    Article Title: Haematopoietic stem cell differentiation promotes the release of prominin-1/CD133-containing membrane vesicles--a role of the endocytic-exocytic pathway

    doi: 10.1002/emmm.201100147

    Figure Lengend Snippet: Provoked differentiation stimulated prominin-1-containing membrane vesicle release A,B. HSPCs pre-cultured either on MSCs or fibronectin-coated dishes for 2 days were incubated in the absence (A) or presence (B) of PMA. After 3 days, conditioned media were subjected to differential centrifugation as in Fig 1A . The resulting pellets were analysed by immunoblotting for prominin-1, actin and α-tubulin. Note that only 1/3 of materials were loaded for the 300 × g fraction. The amount of prominin-1 immunoreactive materials was quantified [ n = 3 (MSCs) and 4 (fibronectin)]. Results are the mean ± SEM. Ratio of prominin-1-immunoreactivity found in the 200,000 × g pellet (vesicles) versus 300 × g (cells) is indicated in bracket. Note that the provoked HSPC differentiation triggered by PMA resulted in an increase of prominin-1 in 200,000 × g pellets, which are devoid of cytoskeletal proteins under both culture conditions.

    Article Snippet: Again, additional investigation is urged to reveal whether prominin-1-CMV modify the biochemistry of the recipient MSCs (Freund et al, ) as it was elegantly proposed upon the direct contact of HSPCs with osteoblasts (Gillette et al, ).

    Techniques: Cell Culture, Incubation, Centrifugation

    Expression of prominin-1 versus either CD61 or CD42b A,B. Flow cytometric analysis revealed phenotypes of HSPCs cultured for 1 week on MSCs for the expression of prominin-1 (CD133) versus either CD61 (A) or CD42b (B). The gates were set according to unstained cells and isotype controls (left panels). The amount of CD133-bright, -dim and -negative cell population is shown as a percentage of the whole CD61- or CD42b-positive population (right panels). C. The percentage of cells harbouring the differentiation markers CD61 and CD42b increased concomitant with the decrease in prominin-1 (CD133) expression, resolving that they are mainly expressed on prominin-1-negative cell population. Data obtained from two independent experiments are shown.

    Journal: EMBO Molecular Medicine

    Article Title: Haematopoietic stem cell differentiation promotes the release of prominin-1/CD133-containing membrane vesicles--a role of the endocytic-exocytic pathway

    doi: 10.1002/emmm.201100147

    Figure Lengend Snippet: Expression of prominin-1 versus either CD61 or CD42b A,B. Flow cytometric analysis revealed phenotypes of HSPCs cultured for 1 week on MSCs for the expression of prominin-1 (CD133) versus either CD61 (A) or CD42b (B). The gates were set according to unstained cells and isotype controls (left panels). The amount of CD133-bright, -dim and -negative cell population is shown as a percentage of the whole CD61- or CD42b-positive population (right panels). C. The percentage of cells harbouring the differentiation markers CD61 and CD42b increased concomitant with the decrease in prominin-1 (CD133) expression, resolving that they are mainly expressed on prominin-1-negative cell population. Data obtained from two independent experiments are shown.

    Article Snippet: Again, additional investigation is urged to reveal whether prominin-1-CMV modify the biochemistry of the recipient MSCs (Freund et al, ) as it was elegantly proposed upon the direct contact of HSPCs with osteoblasts (Gillette et al, ).

    Techniques: Expressing, Flow Cytometry, Cell Culture

    Release of prominin-1-containing membrane vesicles concomitant with haematopoietic stem cell differentiation Differential centrifugation of conditioned HSPC/MSC media recovered after either 4, 9 or 14 days in culture as in Fig 1A . The resulting pellets were analysed by immunoblotting for prominin-1 (arrow) using mAb 80B258 and quantified. Ratio of prominin-1-immunoreactivity found in the 200,000 × g pellet (vesicles) versus the 300 × g pellet (cells) were plotted for the 4, 9 and 14 days of culture (left y -axis; blue). Total number of haematopoietic cells and prominin-1-positive HSPCs present in culture after 4, 9 and 14 days were indicated in the right y -axes, green and red, respectively ( n = 4). The number of prominin-1-positive cells was calculated by multiplying the percentage of prominin-1-positive cells (observed by FACS analysis) with the total number of cells. Note that the general proliferation of prominin-1-positive HSPCs stops after 9 days in culture. Summary of the different conditions. Conditioned media from HSPCs growing on MSCs were recovered after 5 (conditions 4–9 or 9–14) or 10 (conditions 0–9 or 4–14) days in culture. The new medium was added to day 0 (plating day; condition 0–9) or to the 4- (conditions 4–9 and 4–14) or 9- (condition 9–14) days old cultured cells. Differential centrifugation of conditioned media obtained from the four culture conditions described in (C). The resulting pellets were analysed as in (A).

    Journal: EMBO Molecular Medicine

    Article Title: Haematopoietic stem cell differentiation promotes the release of prominin-1/CD133-containing membrane vesicles--a role of the endocytic-exocytic pathway

    doi: 10.1002/emmm.201100147

    Figure Lengend Snippet: Release of prominin-1-containing membrane vesicles concomitant with haematopoietic stem cell differentiation Differential centrifugation of conditioned HSPC/MSC media recovered after either 4, 9 or 14 days in culture as in Fig 1A . The resulting pellets were analysed by immunoblotting for prominin-1 (arrow) using mAb 80B258 and quantified. Ratio of prominin-1-immunoreactivity found in the 200,000 × g pellet (vesicles) versus the 300 × g pellet (cells) were plotted for the 4, 9 and 14 days of culture (left y -axis; blue). Total number of haematopoietic cells and prominin-1-positive HSPCs present in culture after 4, 9 and 14 days were indicated in the right y -axes, green and red, respectively ( n = 4). The number of prominin-1-positive cells was calculated by multiplying the percentage of prominin-1-positive cells (observed by FACS analysis) with the total number of cells. Note that the general proliferation of prominin-1-positive HSPCs stops after 9 days in culture. Summary of the different conditions. Conditioned media from HSPCs growing on MSCs were recovered after 5 (conditions 4–9 or 9–14) or 10 (conditions 0–9 or 4–14) days in culture. The new medium was added to day 0 (plating day; condition 0–9) or to the 4- (conditions 4–9 and 4–14) or 9- (condition 9–14) days old cultured cells. Differential centrifugation of conditioned media obtained from the four culture conditions described in (C). The resulting pellets were analysed as in (A).

    Article Snippet: Again, additional investigation is urged to reveal whether prominin-1-CMV modify the biochemistry of the recipient MSCs (Freund et al, ) as it was elegantly proposed upon the direct contact of HSPCs with osteoblasts (Gillette et al, ).

    Techniques: Cell Differentiation, Centrifugation, FACS, Cell Culture

    Two pools of prominin-1 co-exist in HSPCs A. HSPCs growing for 1 week on MSCs were first cell surface-labelled for prominin-1 prior to PFA fixation and saponin-permeabilization followed by a second round of prominin-1 labelling. The nuclei were stained with Hoechst (blue). The labelled cells were analysed using confocal laser scanning microscopy. A composite of 16 optical sections of HSPCs exhibiting various morphologies as indicated are shown. Note that prominin-1 is not only found at the cell surface (red, arrows) but also in intracellular compartments (green) of the HSPCs. DIC, differential interference contrast. B. Partial co-localization of prominin-1 and endosomal/lysosomal/exosomal marker CD63. PFA-fixed, saponin-permeabilized HSPCs growing on MSCs were double-immunolabelled for prominin-1 (red) and CD63 (green) followed by appropriate secondary antibodies and observed by confocal laser scanning microscopy. Nuclei were visualized with Hoechst (blue). A single optical x – y -plane section revealed that prominin-1 and CD63 partially co-localized in intracellular structures. A high magnification view is shown in the corresponding inset. C,D. Negative staining EM of prominin-1 (C) or CD63 (D) immunogold labelled HSPCs followed by silver enhancement (C) revealed the localization of prominin-1 in either small membrane vesicles present within multivesicular bodies (C, see inset, arrowheads) or microvillar-like structures present at the cell surface (C, arrow). (C) The red box in the left panel demarcates the region displayed at higher magnification in the right panel. The multivesicular bodies were also labelled with CD63 (D). Note that the membranous structures were not well preserved due to saponin-permeabilization (C) prior the labelling procedure. Scale bars, 10 µm [(A) and (B)], 0.5 µm [(C) and (D)].

    Journal: EMBO Molecular Medicine

    Article Title: Haematopoietic stem cell differentiation promotes the release of prominin-1/CD133-containing membrane vesicles--a role of the endocytic-exocytic pathway

    doi: 10.1002/emmm.201100147

    Figure Lengend Snippet: Two pools of prominin-1 co-exist in HSPCs A. HSPCs growing for 1 week on MSCs were first cell surface-labelled for prominin-1 prior to PFA fixation and saponin-permeabilization followed by a second round of prominin-1 labelling. The nuclei were stained with Hoechst (blue). The labelled cells were analysed using confocal laser scanning microscopy. A composite of 16 optical sections of HSPCs exhibiting various morphologies as indicated are shown. Note that prominin-1 is not only found at the cell surface (red, arrows) but also in intracellular compartments (green) of the HSPCs. DIC, differential interference contrast. B. Partial co-localization of prominin-1 and endosomal/lysosomal/exosomal marker CD63. PFA-fixed, saponin-permeabilized HSPCs growing on MSCs were double-immunolabelled for prominin-1 (red) and CD63 (green) followed by appropriate secondary antibodies and observed by confocal laser scanning microscopy. Nuclei were visualized with Hoechst (blue). A single optical x – y -plane section revealed that prominin-1 and CD63 partially co-localized in intracellular structures. A high magnification view is shown in the corresponding inset. C,D. Negative staining EM of prominin-1 (C) or CD63 (D) immunogold labelled HSPCs followed by silver enhancement (C) revealed the localization of prominin-1 in either small membrane vesicles present within multivesicular bodies (C, see inset, arrowheads) or microvillar-like structures present at the cell surface (C, arrow). (C) The red box in the left panel demarcates the region displayed at higher magnification in the right panel. The multivesicular bodies were also labelled with CD63 (D). Note that the membranous structures were not well preserved due to saponin-permeabilization (C) prior the labelling procedure. Scale bars, 10 µm [(A) and (B)], 0.5 µm [(C) and (D)].

    Article Snippet: Again, additional investigation is urged to reveal whether prominin-1-CMV modify the biochemistry of the recipient MSCs (Freund et al, ) as it was elegantly proposed upon the direct contact of HSPCs with osteoblasts (Gillette et al, ).

    Techniques: Staining, Confocal Laser Scanning Microscopy, Marker, Negative Staining

    Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC MSCs) to produce IDO1 (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p

    Journal: Stem Cells Translational Medicine

    Article Title: Serum IFN‐γ Predicts the Therapeutic Effect of Mesenchymal Stem Cells Transplantation in Systemic Lupus Erythematosus Patients

    doi: 10.1002/sctm.17-0002

    Figure Lengend Snippet: Recombinant human IFN‐γ dose dependently enhance indoleamine 2,3‐dioxygenase expression. In vitro, IFN‐γ stimulated umbilical cord‐derived mesenchymal stem cells (UC MSCs) to produce IDO1 (A) as well as IDO2 (B) , both had dose‐dependent manners. High dose IFN‐γ slightly inhibited UC MSCs proliferation (C) , but did not affect MSCs apoptosis (D) . The addition of anti‐IFN‐γ significantly inhibited the production of IDO1 (E) as well as IDO2 (F) by MSCs induced by lupus patients' peripheral blood mononuclear cell. *, p

    Article Snippet: The concentration of IDO1 protein in MSCs (1:400 dilution, Epitomics Technology, Burlingame, CA, http://www.epitomics.com/ ) was also determined.

    Techniques: Recombinant, Expressing, In Vitro, Derivative Assay

    Typical presence of hUCB-MSCs and expression of inflammatory factors following the transplantation of hUCB-MSCs into ICH rats. (A) Three days after the transplantation, hUCB-MSCs were identified by staining with human nuclei antibody (HuNu, red). Confocal micrograph of a brain section show the HuNu-positive cells in peri-infarct regions. The expressions of TNF-α (green) (B), COX-2 (green) (C), CD11b (green) (D), and MPO (green) (E) were detected in the perihemorrhagic regions. The number of TNF-α-positive cells (F), COX-2 positive cells (G), CD11b-reactive microglia (H), and MPO-reactive neutrophils (I) decreased in the hUCB-MSCs-treated group compared with the controls (no significant differences). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM; n=3 per group.

    Journal: Experimental Neurobiology

    Article Title: The Effect of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells in a Collagenase-Induced Intracerebral Hemorrhage Rat Model

    doi: 10.5607/en.2015.24.2.146

    Figure Lengend Snippet: Typical presence of hUCB-MSCs and expression of inflammatory factors following the transplantation of hUCB-MSCs into ICH rats. (A) Three days after the transplantation, hUCB-MSCs were identified by staining with human nuclei antibody (HuNu, red). Confocal micrograph of a brain section show the HuNu-positive cells in peri-infarct regions. The expressions of TNF-α (green) (B), COX-2 (green) (C), CD11b (green) (D), and MPO (green) (E) were detected in the perihemorrhagic regions. The number of TNF-α-positive cells (F), COX-2 positive cells (G), CD11b-reactive microglia (H), and MPO-reactive neutrophils (I) decreased in the hUCB-MSCs-treated group compared with the controls (no significant differences). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM; n=3 per group.

    Article Snippet: Preparation of hUCB-MSCs hUCB-MSCs were provided by MEDIPOST Co., Ltd. (Seoul, Korea).

    Techniques: Expressing, Transplantation Assay, Staining

    Representative results showing neurogenesis, angiogenesis, and apoptosis following the transplantation of hUCB-MSCs into ICH rats. The rats at 4 weeks after transplantation were analyzed for the presence of Tuj1, Laminin, and TUNEL using immunofluorescence staining. (A) Tuj1-positive cells (green) were detected in the SVZ. (B) Laminin-positive cells (green) were present in large numbers in the peri-infarct regions. (C) TUNEL-positive cells (red) were detected in peri-infarct regions. (D) Tuj1-positive cells increased in the hUCB-MSCs-treated group (n=8) compared with the controls (n=10) (not significant). (E) Laminin-positive cells increased significantly the hUCB-MSCs-treated rats (n=8) compared with the controls (n=10). (F) TUNEL-positive cells decreased in the hUCB-MSCs-treated group (n=8) compared with the controls (n=10) (not significant). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM. * p

    Journal: Experimental Neurobiology

    Article Title: The Effect of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells in a Collagenase-Induced Intracerebral Hemorrhage Rat Model

    doi: 10.5607/en.2015.24.2.146

    Figure Lengend Snippet: Representative results showing neurogenesis, angiogenesis, and apoptosis following the transplantation of hUCB-MSCs into ICH rats. The rats at 4 weeks after transplantation were analyzed for the presence of Tuj1, Laminin, and TUNEL using immunofluorescence staining. (A) Tuj1-positive cells (green) were detected in the SVZ. (B) Laminin-positive cells (green) were present in large numbers in the peri-infarct regions. (C) TUNEL-positive cells (red) were detected in peri-infarct regions. (D) Tuj1-positive cells increased in the hUCB-MSCs-treated group (n=8) compared with the controls (n=10) (not significant). (E) Laminin-positive cells increased significantly the hUCB-MSCs-treated rats (n=8) compared with the controls (n=10). (F) TUNEL-positive cells decreased in the hUCB-MSCs-treated group (n=8) compared with the controls (n=10) (not significant). Nuclei were counterstained with DAPI (blue). Values are shown as mean and SEM. * p

    Article Snippet: Preparation of hUCB-MSCs hUCB-MSCs were provided by MEDIPOST Co., Ltd. (Seoul, Korea).

    Techniques: Transplantation Assay, TUNEL Assay, Immunofluorescence, Staining

    Transplantation of hUCB-MSCs reduces lesion volume and improves the behavioral recovery of ICH rats. (A) Schematic timeline of the experimental procedures. MSCs indicates mesenchymal stem cells; Sacr, sacrifice; IF, immunofluorescence staining; WB, western blot; and BT, behavior test. (B) H E staining of coronal sections for characterization of unilateral brain injury at 30 days after ICH. (C) The volume of lesion in the hUCB-MSCs-treated rats (n=8) was significantly smaller than in the controls (n=10) at 4 weeks after transplantation. ** p

    Journal: Experimental Neurobiology

    Article Title: The Effect of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells in a Collagenase-Induced Intracerebral Hemorrhage Rat Model

    doi: 10.5607/en.2015.24.2.146

    Figure Lengend Snippet: Transplantation of hUCB-MSCs reduces lesion volume and improves the behavioral recovery of ICH rats. (A) Schematic timeline of the experimental procedures. MSCs indicates mesenchymal stem cells; Sacr, sacrifice; IF, immunofluorescence staining; WB, western blot; and BT, behavior test. (B) H E staining of coronal sections for characterization of unilateral brain injury at 30 days after ICH. (C) The volume of lesion in the hUCB-MSCs-treated rats (n=8) was significantly smaller than in the controls (n=10) at 4 weeks after transplantation. ** p

    Article Snippet: Preparation of hUCB-MSCs hUCB-MSCs were provided by MEDIPOST Co., Ltd. (Seoul, Korea).

    Techniques: Transplantation Assay, Immunofluorescence, Staining, Western Blot

    RT-PCR and western blot analysis of inflammatory factors. The mRNA and protein levels were detected in whole brain tissue of ICH rats at 3 days after transplantation. (A) mRNA levels of inflammatory factors (TNF-α, COX-2, CD11b, and MPO) by RT-PCR. (B~E) Quantification of the mRNA expression of inflammatory factors in the hUCB-MSCs-treated group compared with the controls. (F) Representative western blots of the four inflammatory factors (G~J). Quantification (protein expression) of inflammatory factors in the hUCB-MSCs-treated group compared with the controls. mRNA and protein levels of inflammatory factors were lower in the hUCB-MSCs-treated group than in the controls (not significant). Densitometric analysis was performed using the Image J software by normalizing to GAPDH for RT-PCR and β-actin for western blot as the loading control; n=3 per group.

    Journal: Experimental Neurobiology

    Article Title: The Effect of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells in a Collagenase-Induced Intracerebral Hemorrhage Rat Model

    doi: 10.5607/en.2015.24.2.146

    Figure Lengend Snippet: RT-PCR and western blot analysis of inflammatory factors. The mRNA and protein levels were detected in whole brain tissue of ICH rats at 3 days after transplantation. (A) mRNA levels of inflammatory factors (TNF-α, COX-2, CD11b, and MPO) by RT-PCR. (B~E) Quantification of the mRNA expression of inflammatory factors in the hUCB-MSCs-treated group compared with the controls. (F) Representative western blots of the four inflammatory factors (G~J). Quantification (protein expression) of inflammatory factors in the hUCB-MSCs-treated group compared with the controls. mRNA and protein levels of inflammatory factors were lower in the hUCB-MSCs-treated group than in the controls (not significant). Densitometric analysis was performed using the Image J software by normalizing to GAPDH for RT-PCR and β-actin for western blot as the loading control; n=3 per group.

    Article Snippet: Preparation of hUCB-MSCs hUCB-MSCs were provided by MEDIPOST Co., Ltd. (Seoul, Korea).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Transplantation Assay, Expressing, Software