Journal: Stem Cells International

Article Title: Acute Leukemia Induces Senescence and Impaired Osteogenic Differentiation in Mesenchymal Stem Cells Endowing Leukemic Cells with Functional Advantages

doi: 10.1155/2019/3864948

Figure Lengend Snippet: Adhesion molecule expression in MSC from the LN and the REH-CM LN. MSC were cocultured with REH cells and REH-CM for three days. Flow cytometry analysis of (a) CD106, (b) CD54, (c) CD49e, (d) CD184, and (e) SDF-1 were measured as indicated. Results are expressed as the median fluorescence intensity (MFI) from two independent experiments done in triplicates ( p values: Student t -tests; ns: nonsignificant, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001).

Article Snippet: MSC from the different culture conditions were trypsinized, washed twice with PBS 1x, and stained with different monoclonal antibodies for flow cytometry analysis: PE-conjugated mouse anti-human CD49e (clone IIA1, BD Pharmingen, San Jose, CA, USA), APC-conjugated mouse anti-human CD49d (clone 9F10, BD Pharmingen, San Jose, CA, USA), APC-conjugated mouse anti-human CD54 (clone REA266, Miltenyi Biotec, Auburn, CA, USA), PE-conjugated CD106 (VCAM-1) (clone REA269, Miltenyi Biotec, Auburn, CA, USA), FITC-conjugated mouse anti-human CD44 (clone MEM-85, Invitrogen, Frederick, MD, USA), and PE-conjugated CD184 (CXCR4) (clone 12G5, Miltenyi Biotec, Auburn, CA, USA).

Techniques: Expressing, Flow Cytometry, Fluorescence

Journal: Stem Cells International

Article Title: Acute Leukemia Induces Senescence and Impaired Osteogenic Differentiation in Mesenchymal Stem Cells Endowing Leukemic Cells with Functional Advantages

doi: 10.1155/2019/3864948

Figure Lengend Snippet: REH cell evaluation after MSC coculture. (a) REH cells were cocultured with MSC for three days after which cell proliferation was assessed. Proliferation capacity was measured by cell staining with CFSE (left panel). Cells were synchronized by serum starvation (FBS removal for two days). Percentage of synchronized REH cells (black bars), monoculture (white bars), and coculture (grey bars) at the corresponding number of cell division (0, 1, and 2) are shown (right panel). (b) Adhesion molecule expression in REH cells cultured in LN conditions for three days. Median fluorescence intensity (MFI) of CD49d, CD49e, CD54, CD44, and CD184 is shown. (c) REH cell migration capacity towards SDF-1 (100 ng/mL) was determined in a transwell insert with a 5 μ m pore membrane. REH cells in monoculture and coculture with MSC were allowed to migrate for 24 h, after which cells in the lower chamber were harvested and counted by flow cytometry. The percentage of migration was calculated considering the total input of REH cells. Data were obtained from two independent experiments done in triplicates ( p values: Student t -test; ns: nonsignificant, ∗ p < 0.05 and ∗∗ p < 0.01).

Article Snippet: MSC from the different culture conditions were trypsinized, washed twice with PBS 1x, and stained with different monoclonal antibodies for flow cytometry analysis: PE-conjugated mouse anti-human CD49e (clone IIA1, BD Pharmingen, San Jose, CA, USA), APC-conjugated mouse anti-human CD49d (clone 9F10, BD Pharmingen, San Jose, CA, USA), APC-conjugated mouse anti-human CD54 (clone REA266, Miltenyi Biotec, Auburn, CA, USA), PE-conjugated CD106 (VCAM-1) (clone REA269, Miltenyi Biotec, Auburn, CA, USA), FITC-conjugated mouse anti-human CD44 (clone MEM-85, Invitrogen, Frederick, MD, USA), and PE-conjugated CD184 (CXCR4) (clone 12G5, Miltenyi Biotec, Auburn, CA, USA).

Techniques: Staining, Expressing, Cell Culture, Fluorescence, Migration, Membrane, Flow Cytometry

Journal: Stem Cells International

Article Title: Migration, Proliferation, and Differentiation of Cord Blood Mesenchymal Stromal Cells Treated with Histone Deacetylase Inhibitor Valproic Acid

doi: 10.1155/2014/610495

Figure Lengend Snippet: Primers used for gel-based RT-PCR analysis.

Article Snippet: The surface expression of CXCR4 and CXCR7 on CB-derived MSC was examined using PE-conjugated antihuman CD184 (CXCR4, clone 12G5, BD Biosciences Mississauga, ON, Canada) and PE-conjugated antihuman CXCR7 (clone 11G8, R&D Systems, Minneapolis, MN, USA).

Techniques: Sequencing

Journal: Stem Cells International

Article Title: Migration, Proliferation, and Differentiation of Cord Blood Mesenchymal Stromal Cells Treated with Histone Deacetylase Inhibitor Valproic Acid

doi: 10.1155/2014/610495

Figure Lengend Snippet: VPA increases CXCR4 and CXCR7 gene and protein expression. CB-derived MSC were treated with 0 (control), 1, 5, or 10 mM of VPA for 3 or 6 h. (a) Expression of CXCR4 and CXCR7 mRNAs was evaluated by real-time quantitative RT-PCR using 18S mRNA as internal calibrator. The data shown are based on two independent experiments. * P < 0.05 indicates statistically significant difference relative to control. (b) Surface expression of CXCR4 and CXCR7 was not affected by VPA treatment as determined by flow cytometry. The black line represents isotype control, the red line is for CXCR4, and the green line is for CXCR7. (c) Western blot of total CXCR4 and CXCR7 protein using β -actin as loading control. The numbers at the bottom of the gels represent the fold-increase in expression after VPA treatment relative to control. The data is representative of three independent experiments.

Article Snippet: The surface expression of CXCR4 and CXCR7 on CB-derived MSC was examined using PE-conjugated antihuman CD184 (CXCR4, clone 12G5, BD Biosciences Mississauga, ON, Canada) and PE-conjugated antihuman CXCR7 (clone 11G8, R&D Systems, Minneapolis, MN, USA).

Techniques: Expressing, Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Western Blot

Journal: Stem Cells International

Article Title: Migration, Proliferation, and Differentiation of Cord Blood Mesenchymal Stromal Cells Treated with Histone Deacetylase Inhibitor Valproic Acid

doi: 10.1155/2014/610495

Figure Lengend Snippet: VPA enhances migration of CB MSC towards a low SDF-1 gradient which is inhibited by CXCR4 and CXCR7 antagonists. CB-derived MSC were preincubated with or without 5 mM VPA for 3 h and allowed to migrate across Matrigel towards a low (20 ng/mL) or high (100 ng/mL) SDF-1 gradient. Cells were also incubated with CXCR4 antagonist AMD3100, CXCR7 antagonist CCX733, or the inactive CXCR7 antagonist CCX226. The data are based on two independent experiments; P ≤ 0.001 indicates statistically significant difference.

Article Snippet: The surface expression of CXCR4 and CXCR7 on CB-derived MSC was examined using PE-conjugated antihuman CD184 (CXCR4, clone 12G5, BD Biosciences Mississauga, ON, Canada) and PE-conjugated antihuman CXCR7 (clone 11G8, R&D Systems, Minneapolis, MN, USA).

Techniques: Migration, Derivative Assay, Incubation

Journal: Heliyon

Article Title: Chronic treatment with TNF-α, alone and in combination with Takinib, SB203580 and metformin induce cell death in breast cancer

doi: 10.1016/j.heliyon.2023.e21060

Figure Lengend Snippet: CXCR4+ population after treating cells with TNF-α for one week or its combination with SB203580 or Metformin or Takinib for 24 and 48 h compared to control (untreated) cells. Percentage of CXCR4+ population in MCF-7 cells (A) and in MDA-MB-231 cells (B). Example of the CXCR4+ population in MCF-7 (C) and MDA-MB-231 cells (D) . The data represent n = 3 (Mean ± SD). P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: FITC-conjugated anti-CD24 antibody (Clone M1/69), PE -conjugated anti-CD44 antibody, (Clone IM7) and PE-Cyanine5-conjugated anti-CD184 (CXCR4) antibody (Clone 12G5) were obtained from eBioscience.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Dual Targeting of Stromal Cell Support and Leukemic Cell Growth by a Peptidic PKC Inhibitor Shows Effectiveness against B-ALL

doi: 10.3390/ijms21103705

Figure Lengend Snippet: Cell to cell interaction promotes changes in the expression profile of different adhesion molecules in MSC and B-ALL cell populations. ( A ) A representative experiment showing the expression of VLA-4, VLA-5, VCAM, ICAM-1, CXCR4, and CD44 by flow cytometry in MSC alone or in co-cultures with B-ALL cells stablished for 6 h. ( B ) Unattached B-ALL cells and B-ALL cells recovered from the co-culture were labelled for assessing the expression of the indicated molecules. Data are expressed as mean of MFI ± SEM obtained from three independent experiments.

Article Snippet: The following monoclonal antibodies were used for staining MSC and B-ALL cells, as previously described [ ]: PE mouse anti-human CD49e (clone IIA1, BD Pharmingen, San Jose, CA, USA), APC mouse anti-human CD49d (clone 9F10, BD Pharmingen) or BV711 mouse anti-human CD49d (clone 9F10, BD Horizon) APC mouse anti-human CD54 (clone REA266, Miltenyi Biotec, Auburn, CA, USA), PE mouse anti-human CD106 (VCAM-1) (clone REA269, Miltenyi Biotec) or BV605 mouse anti-human CD106 (Clone 51-10C9, BD Horizon), FITC mouse anti-human CD44 (clone G44-26, BD Pharmingen) or APC mouse anti-human CD44 (clone G44-26) and PE CD184 (CXCR4) (clone 12G5, Miltenyi Biotec).

Techniques: Expressing, Flow Cytometry, Co-Culture Assay

Journal: International Journal of Molecular Sciences

Article Title: Dual Targeting of Stromal Cell Support and Leukemic Cell Growth by a Peptidic PKC Inhibitor Shows Effectiveness against B-ALL

doi: 10.3390/ijms21103705

Figure Lengend Snippet: HKPS treatment reduces the expression of molecules involved in the MSC and B-ALL interaction. ( A ) MSC were pre-treated for 2 h with HKPS (40 μM), STAU (0.5 μM) or vehicle (DMSO 0.4%) and then co-cultures with B-ALL cells were stablished for 6 h. A representative experiment showing the expression of VLA-4, VLA-5, VCAM, ICAM-1, CXCR4, and CD44 evaluated by flow cytometry in CD105+ cell population. MSC alone were used as control. ( B ) Unattached B-ALL cells and B-ALL cells recovered after trypsinization from the co-cultures with MSC were labelled for assessing the expression of the indicated molecules in the CD19+ cell population. Data are expressed as mean of MFI ± SEM obtained from two independent experiments.

Article Snippet: The following monoclonal antibodies were used for staining MSC and B-ALL cells, as previously described [ ]: PE mouse anti-human CD49e (clone IIA1, BD Pharmingen, San Jose, CA, USA), APC mouse anti-human CD49d (clone 9F10, BD Pharmingen) or BV711 mouse anti-human CD49d (clone 9F10, BD Horizon) APC mouse anti-human CD54 (clone REA266, Miltenyi Biotec, Auburn, CA, USA), PE mouse anti-human CD106 (VCAM-1) (clone REA269, Miltenyi Biotec) or BV605 mouse anti-human CD106 (Clone 51-10C9, BD Horizon), FITC mouse anti-human CD44 (clone G44-26, BD Pharmingen) or APC mouse anti-human CD44 (clone G44-26) and PE CD184 (CXCR4) (clone 12G5, Miltenyi Biotec).

Techniques: Expressing, Flow Cytometry