Journal: PLoS ONE

Article Title: Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro

doi: 10.1371/journal.pone.0132480

Figure Lengend Snippet: There were no significant differences between the control group and the NDIFE hADMSC groups at each time point (one-way analysis of variance, P > 0.05). Error bars represent the mean ± standard deviation (n = 3). The ferritin transgene did not affect the proliferation rate of hADMSCs.

Article Snippet: Second-passage hADMSCs (cat. # C0013; Biowit Technologies, Shenzhen, China; obtained directly from the company) were seeded, cultured, and propagated in complete medium (high-glucose DMEM supplemented with 10% FBS, 10 mM N -2-hydroxyethylpiperazine- N -2-ethane sulfonic acid, 2 mM l -glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin) in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Control, Standard Deviation

Journal: PLoS ONE

Article Title: Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro

doi: 10.1371/journal.pone.0132480

Figure Lengend Snippet: After neural differentiation, neural-differentiation-inducible ferritin-expressing (NDIFE) human adipose tissue-derived mesenchymal stem cells (hADMSCs) began to express neural cell-specific markers. The scale bar is 50 μm. SYN1: synapsin I, NSE: neuron-specific enolase, FTH1: ferritin heavy chain 1, GFAP: glial fibrillary acidic protein, MBP: myelin basic protein, DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Second-passage hADMSCs (cat. # C0013; Biowit Technologies, Shenzhen, China; obtained directly from the company) were seeded, cultured, and propagated in complete medium (high-glucose DMEM supplemented with 10% FBS, 10 mM N -2-hydroxyethylpiperazine- N -2-ethane sulfonic acid, 2 mM l -glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin) in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Expressing, Derivative Assay

Journal: PLoS ONE

Article Title: Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro

doi: 10.1371/journal.pone.0132480

Figure Lengend Snippet: (A) Western blot analysis of ferritin expression in neural-differentiation-inducible ferritin-expressing (NDIFE) and neurally differentiated (ND)-NDIFE human adipose tissue-derived mesenchymal stem cells (hADMSCs). Ferritin expression was significantly higher in ND-NDIFE hADMSCs than in the corresponding control NDIFE hADMSCs. Error bars represent the mean ± standard deviation (n = 3); * P < 0.05. (B) A photomicrograph showing the immunofluorescent staining of ferritin in NDIFE and ND-NDIFE hADMSCs. Ferritin expression (red fluorescence) was significantly stronger in ND-NDIFE hADMSCs than in the corresponding control NDIFE hADMSCs. The scale bar is 50 μm. GFP: green fluorescent protein, DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Second-passage hADMSCs (cat. # C0013; Biowit Technologies, Shenzhen, China; obtained directly from the company) were seeded, cultured, and propagated in complete medium (high-glucose DMEM supplemented with 10% FBS, 10 mM N -2-hydroxyethylpiperazine- N -2-ethane sulfonic acid, 2 mM l -glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin) in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Western Blot, Expressing, Derivative Assay, Control, Standard Deviation, Staining, Fluorescence

Journal: PLoS ONE

Article Title: Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro

doi: 10.1371/journal.pone.0132480

Figure Lengend Snippet: (A) A photomicrograph showing Prussian blue iron staining of intracellular iron in neural-differentiation-inducible ferritin-expressing (NDIFE) and neurally differentiated (ND)-NDIFE human adipose tissue-derived mesenchymal stem cells (hADMSCs). Many disperse cytoplasmic deposits (blue granules) of accumulated intracellular iron were observed in NDIFE hADMSCs, whereas large, dense cytoplasmic deposits appeared in the corresponding control ND-NDIFE hADMSCs. The scale bar is 50 μm. (B) Inductively coupled plasma mass spectrometry (ICP-MS) analysis of the intracellular iron content in NDIFE and ND-NDIFE hADMSCs. The intracellular iron content in ND-NDIFE hADMSCs was significantly higher than that in the corresponding undifferentiated NDIFE hADMSCs. Error bars represent the mean ± standard deviation (n = 3; * P < 0.05).

Article Snippet: Second-passage hADMSCs (cat. # C0013; Biowit Technologies, Shenzhen, China; obtained directly from the company) were seeded, cultured, and propagated in complete medium (high-glucose DMEM supplemented with 10% FBS, 10 mM N -2-hydroxyethylpiperazine- N -2-ethane sulfonic acid, 2 mM l -glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin) in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Staining, Expressing, Derivative Assay, Control, Clinical Proteomics, Mass Spectrometry, Standard Deviation

Journal: PLoS ONE

Article Title: Use of Ferritin Expression, Regulated by Neural Cell-Specific Promoters in Human Adipose Tissue-Derived Mesenchymal Stem Cells, to Monitor Differentiation with Magnetic Resonance Imaging In Vitro

doi: 10.1371/journal.pone.0132480

Figure Lengend Snippet: A T2-weighted image from magnetic resonance imaging (MRI) of agarose phantoms of neural-differentiation-inducible ferritin-expressing (NDIFE) and neurally differentiated (ND)-NDIFE human adipose tissue-derived mesenchymal stem cells (hADMSCs). The R2 relaxation rate of ND-NDIFE hADMSCs was significantly higher than that of the corresponding undifferentiated NDIFE hADMSCs, resulting in notable hypointensity in T2-weighted images of ND-NDIFE hADMSCs. Error bars represent the mean ± standard deviation (n = 4; * P < 0.05).

Article Snippet: Second-passage hADMSCs (cat. # C0013; Biowit Technologies, Shenzhen, China; obtained directly from the company) were seeded, cultured, and propagated in complete medium (high-glucose DMEM supplemented with 10% FBS, 10 mM N -2-hydroxyethylpiperazine- N -2-ethane sulfonic acid, 2 mM l -glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin) in a humidified atmosphere containing 5% CO 2 at 37°C.

Techniques: Magnetic Resonance Imaging, Expressing, Derivative Assay, Standard Deviation

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: Identification of a model that induced hAdMSCs to adipocytes differentiation. (A) Expression of adipogenic marker proteins in different stages (D0, D3, D6, D9, and D12) was detected by Western blot, and GAPDH was used as a loading control in each sample. Expression of (B) C/EBPα, (C) PPARγ2, (D) AdipoQ, (E) FABP4, and (F) LPL were quantified using Image J software. The levels of lipogenesis enzymes, such as (G) ACC and (H) FAS were detected by spectrophotometry. (I) Expression of HCG11 in different stages (D0, D3, D6, D9, and D12) was detected by RT-qPCR. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. ** P < 0.01 and *** P < 0.001 versus D0. ACC: acetyl coenzyme A carboxylase; AdipoQ: adiponectin; C/EBPα: CCAAT-enhancer-binding protein α; FABP4: fatty acid-binding protein 4; FAS: fatty acid synthase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hAdMSCs: human adipose-derived mesenchymal stem cells; HCG11: human leukocyte antigen complex group 11; LPL: lipoprotein lipase; PPARγ: peroxisome proliferator-activated receptor gamma; RT-qPCR: real-time quantitative polymerase chain reaction.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Expressing, Marker, Western Blot, Control, Software, Spectrophotometry, Quantitative RT-PCR, Binding Assay, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: Differentiation of adipocytes overexpressing lncRNA HCG11. (A) Expression of HCG11 in different stages (D0, D6, and D12) of hAdMSCs transfected with pcDNA-HCG11 was detected by RT-qPCR. (B) Expression of adipogenic marker protein was detected by Western blot, and GAPDH was used as a loading control in each sample. The expression of (C) C/EBPα, (D) PPARγ2, (E) AdipoQ, (F) FABP4, and (G) LPL in different stages (D0, D6, and D12) of hAdMSCs transfected with pcDNA-HCG11 and control was quantified using Image J software. The levels of lipogenesis enzymes such as (H) ACC and (I) FAS were detected by spectrophotometry. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. * P < 0.05 and ** P < 0.01 versus control; # P < 0.05 and ## P < 0.01 versus vector. ACC: acetyl coenzyme A carboxylase; AdipoQ: adiponectin; C/EBPα: CCAAT-enhancer-binding protein α; FABP4: fatty acid-binding protein 4; FAS: fatty acid synthase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hAdMSCs: human adipose-derived mesenchymal stem cells; HCG11: human leukocyte antigen complex group 11; LPL: lipoprotein lipase; PPARγ: peroxisome proliferator-activated receptor gamma; RT-qPCR: real-time quantitative polymerase chain reaction.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Marker, Western Blot, Control, Software, Spectrophotometry, Plasmid Preparation, Binding Assay, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: Differentiation of adipocytes interfering with sh-HCG11. (A) Expression of HCG11 in different stages (D0, D6, and D12) hAdMSCs transfected with sh-HCG11 was detected by RT-qPCR. (B) Expression of adipogenic marker protein such as (C) C/EBPα, (D) PPARγ2, (E) AdipoQ, (F) FABP4, and (G) LPL was detected by Western blot and quantified using Image J software. The levels of lipogenesis enzymes such as (H) ACC and (I) FAS were detected by spectrophotometry. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. * P < 0.05 and ** P < 0.01 versus control; # P < 0.05 and ## P < 0.01 versus vector. ACC: acetyl coenzyme A carboxylase; AdipoQ: adiponectin; C/EBPα: CCAAT-enhancer-binding protein α; FABP4: fatty acid-binding protein 4; FAS: fatty acid synthase; hAdMSCs: human adipose-derived mesenchymal stem cells; HCG11: human leukocyte antigen complex group 11; LPL: lipoprotein lipase; PPARγ: peroxisome proliferator-activated receptor gamma; RT-qPCR: real-time quantitative polymerase chain reaction.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Expressing, Transfection, Quantitative RT-PCR, Marker, Western Blot, Software, Spectrophotometry, Control, Plasmid Preparation, Binding Assay, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: HCG11 directly targeted to miR-204-5p. (A) Online database StarBase showed the binding sites of HCG11 and miR-204-5p. (B) The luciferase reporter gene assay was performed in HEK-293T cells to validate the binding of HCG11 and miR-204-5p. Firefly and Renilla luciferase activities were determined. (C) HEK-293T cells were transfected with biotinylated miR-204-5p (Bio-204-5p-wt) or its mutant form (Bio-204-5p-mut), and then a biotin-based pull-down assay was performed to detect HCG11 expression and normalized to a biotinylated mimic control (Bio-NC). (D) Expression of miR-204-5p in hAdMSCs transfected with pcDNA-HCG11 or sh-HCG11 was detected by RT-qPCR. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. ** P < 0.01 and *** P < 0.001 versus control; # P < 0.05 versus scramble. hAdMSCs: human adipose-derived mesenchymal stem cells; HCG11: human leukocyte antigen complex group 11; HEK-293T: human embryonic kidney 293T; RT-qPCR: real-time quantitative polymerase chain reaction.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Transfection, Mutagenesis, Pull Down Assay, Expressing, Control, Quantitative RT-PCR, Derivative Assay, Real-time Polymerase Chain Reaction

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: Overexpression of miR-204-5p promoted cell proliferation and adipocytes differentiation in hAdMSCs. The hAdMSCs were transfected with NC mimic (20 nM), miR-204-5p mimic (20 nM), NC inhibitor (20 nM), and miR-204-5p inhibitor (20 nM) for 48 h. (A) Expression of miR-204-5p was detected by RT-qPCR. (B) The expression of adipogenic marker protein and inflammatory factor was detected by Western blot. Expression of (C) C/EBPα, (D) PPARγ2, (E) AdipoQ, (F) FABP4, (G) LPL, (H) IL-6, and (I) TNF-α was quantified using Image J software. The levels of lipogenesis enzymes such as ACC (J) and FAS (K) were detected by spectrophotometry. (L) Cell proliferation ability was tested by CCK-8. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. * P < 0.05 and ** P < 0.01 versus control. ACC: acetyl coenzyme A carboxylase; AdipoQ: adiponectin; C/EBPα: CCAAT-enhancer-binding protein α; CCK: cell counting kit; FABP4: fatty acid-binding protein 4; FAS: fatty acid synthase; hAdMSCs: human adipose-derived mesenchymal stem cells; IL-6: interleukin-6; LPL: lipoprotein lipase; NC: normal control; PPARγ: peroxisome proliferator-activated receptor gamma; TNF-α: tumor necrosis factor-alpha.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Over Expression, Transfection, Expressing, Quantitative RT-PCR, Marker, Western Blot, Software, Spectrophotometry, CCK-8 Assay, Control, Binding Assay, Cell Counting, Derivative Assay

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: MiR-204-5p directly targeted SIRT1. (A) Online database StarBase showed the sequence alignment between miR-204-5p and SIRT1. (B) The luciferase reporter gene assay was performed in HEK-293T cells to validate the binding of miR-204-5p and SIRT1. Firefly and Renilla luciferase activities were determined. (C) Expression of SIRT1 in hAdMSCs transfected with miR-204-5p mimic, inhibitor, and their control was detected by Western blotting. (D) The expression of adipogenic marker protein and inflammatory factor was detected by Western blot. Expression of (E) C/EBPα, (F) PPARγ2, (G) AdipoQ, (H) FABP4, (I) LPL, (J) IL-6, and (K) TNF-α in hAdMSCs transfected with miR-204-5p mimic, Res, and pcDNA-SIRT1. The levels of lipogenesis enzymes such as ACC (L) and FAS (M) were detected by spectrophotometry. (N) Cell proliferation ability was tested by CCK-8. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. * P < 0.05 and ** P < 0.01 versus control. ACC: acetyl coenzyme A carboxylase; AdipoQ: adiponectin; C/EBPα: CCAAT-enhancer-binding protein α; CCK: cell counting kit; FABP4: fatty acid-binding protein 4; FAS: fatty acid synthase; hAdMSCs: human adipose-derived mesenchymal stem cells; IL-6: interleukin-6; LPL: lipoprotein lipase; PPARγ: peroxisome proliferator-activated receptor gamma; TNF-α: tumor necrosis factor alpha.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Sequencing, Luciferase, Reporter Gene Assay, Binding Assay, Expressing, Transfection, Control, Western Blot, Marker, Spectrophotometry, CCK-8 Assay, Cell Counting, Derivative Assay

Journal: Cell Transplantation

Article Title: LncRNA HCG11 Inhibits Adipocyte Differentiation in Human Adipose-Derived Mesenchymal Stem Cells by Sponging miR-204-5p to Upregulate SIRT1

doi: 10.1177/0963689720968090

Figure Lengend Snippet: Overexpression of miR-204-5p reversed the effect of HCG11 on hAdMSCs. (A) The expression of adipogenic marker protein and inflammatory factor was detected by Western blot. Expression of (B) C/EBPα, (C) PPARγ2, (D) AdipoQ, (E) FABP4, (F) LPL, (G) IL-6, and (H) TNF-α in hAdMSCs transfected with miR-204-5p mimic, pcDNA-HCG11, and their control was quantified using Image J software. The levels of lipogenesis enzymes such as (I) ACC and (J) FAS were detected by spectrophotometry. (K) Cell proliferation ability was tested by CCK-8. Statistical significance was determined using an independent sample t -test. Values were expressed as mean ± SEM, n = 3. * P < 0.05 and ** P < 0.01 versus control. ACC: acetyl coenzyme A carboxylase; AdipoQ: adiponectin; C/EBPα: CCAAT-enhancer-binding protein α; CCK: cell counting kit; FABP4: fatty acid-binding protein 4; FAS: fatty acid synthase; hAdMSCs: human adipose-derived mesenchymal stem cells; HCG11: human leukocyte antigen complex group 11; IL-6: interleukin-6; LPL: lipoprotein lipase; PPARγ: peroxisome proliferator-activated receptor gamma; TNF-α: tumor necrosis factor alpha.

Article Snippet: Human embryonic kidney 293T (HEK-293T) cells (Shanghai, China) and hAdMSCs (Procell, Wuhan, Hubei, China) were resuspended in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific, Waltham, MA, USA) mixture supplemented with 10% fetal bovine serum (FBS, GIBCO, Carlsbad, CA, USA), 100 U/ml penicillin, and 100 μg/ml streptomycin (Thermo Fisher Scientific) in a humidified incubator containing 5% carbon dioxide (CO 2 ) at 37 °C. hAdMSCs at passage five were grown for 2 days, followed by differentiation into adipocyte.

Techniques: Over Expression, Expressing, Marker, Western Blot, Transfection, Control, Software, Spectrophotometry, CCK-8 Assay, Binding Assay, Cell Counting, Derivative Assay

Journal: Applied Microbiology and Biotechnology

Article Title: Mesenchymal and induced pluripotent stem cell–based therapeutics: a comparison

doi: 10.1007/s00253-023-12583-4

Figure Lengend Snippet: Overview of approved hMSC-based therapeutics and hiPSC-based therapeutics in phase 2 and 3 clinical trials

Article Snippet: AlloSource , AlloStem ® , Allogeneic hAD-MSCs , Bone regeneration , Approved , USA.

Techniques:

Journal: Bioactive Materials

Article Title: Exosomes derived from differentiated human ADMSC with the Schwann cell phenotype modulate peripheral nerve-related cellular functions

doi: 10.1016/j.bioactmat.2021.11.022

Figure Lengend Snippet: Characterization and comparison of hADMSCs with and without differentiation into SCs. (A) Schematic of the differentiation protocol of hADMSCs into SC-like cells. (B) Optical microscope and IF images of hADMSCs and hADMSC-SCs. (Optical microscope image scale bar: 250 μm, IF image scale bar: 100 μm). (C) Representative images of a human growth factor antibody array for supernatant culture media of hADMSCs and hADMSC-SCs. (D) Semi-quantitative analysis of the secreted growth factors with statistical difference. Relative expression of each targeted growth factor was presented as normalized to the positive control on the same array and relative to the hADMSC control group (n = 4, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Primary hADMSCs (Lonza, USA) before passage 6 were cultured in Dulbecco's Modified Eagle's Medium/F12 (DMEM/F12, HyClone, USA) with 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin/streptomycin (P/S, Invitrogen, USA) at 37 °C and 5% CO 2 .

Techniques: Comparison, Microscopy, Ab Array, Expressing, Positive Control, Control

Journal: Bioactive Materials

Article Title: Exosomes derived from differentiated human ADMSC with the Schwann cell phenotype modulate peripheral nerve-related cellular functions

doi: 10.1016/j.bioactmat.2021.11.022

Figure Lengend Snippet: Characterizations of exosomes derived from hADMSCs and hADMSC-SCs. (A) Schematic of the protocol for exosome isolation from cell culture media. (B) Size distribution of uExo and dExo was obtained from NTA test, inset showing representative exosome images captured from the NTA video frames. (C) AFM images of uExo and dExo (scale bar 250 nm). (D) TEM images of uExo and dExo (scale bar 100 nm). (E) Proteins from each centrifugal rate (1k × g : 1000 × g , 10k × g:10,000 × g , and 100k × g: 100,000 × g ) were analyzed by western blot for the presence of exosome-associated proteins CD9, CD63, TSG101, and Alix.

Article Snippet: Primary hADMSCs (Lonza, USA) before passage 6 were cultured in Dulbecco's Modified Eagle's Medium/F12 (DMEM/F12, HyClone, USA) with 10% fetal bovine serum (FBS, Gibco, USA) and 1% penicillin/streptomycin (P/S, Invitrogen, USA) at 37 °C and 5% CO 2 .

Techniques: Derivative Assay, Isolation, Cell Culture, Western Blot

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: ZD ameliorates cell injury induced by LPS and H2O2 in hADMSCs. (A, B) Measurement of cell viability and apoptotic ratio after different doses of ZD (gray column) with or without LPS/H2O2 coincubation (dark gray column). A proven stem cell protective agent, edaravone (light gray column), was used as the positive control. (C, D) Activities of caspases 3/7 or caspase 8 from cell lysates after LPS/H2O2 incubation with or without ZD cotreatment. (E, F) Secreted TNF-a and IL-6 level changes after LPS/H2O2 incubation with or without ZD cotreatment. (G, H) Intracellular production of ROS was detected by fluorescence probe DCFH-DA (200x) after LPS/H2O2 incubation with or without ZD cotreatment. (I) Changes of ratio between glutathione (GSH) to oxidized glutathione (GSSG) and (J) Western blot for the expression of CAT and SOD1 after LPS/H2O2 incubation with or without ZD cotreatment. The treatment duration is 24 h. *p < 0.05, **p < 0.01, ***p < 0.001 mean significant changes between control and treatments, respectively; ###p < 0.001 means significant changes between indicated treatment with LPS/H2O2 incubation group. L + H, LPS/H2O2; Eda, edaravone; v-ZD, vehicle-ZD treatment; Ctrl, control; ZD, zeaxanthin dipalmitate; hADMSCs, human adipose-derived mesenchymal stem cells; LPS, lipopolysaccharide; TNF-α, tumor necrosis factor-α; IL-6, interleukin-6; DCFH-DA, 2′,7′-dichlorofluorescin diacetate; CAT, catalase; SOD1, superoxide dismutase 1; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; ROS, reactive oxygen species.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Positive Control, Incubation, Fluorescence, Western Blot, Expressing, Control, Derivative Assay

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: Therapeutic effects of transplanted hADMSCs in a murine ALF model were significantly increased by ZD pretreatment and impaired by stable knockdown of cellular miR-210 expression, respectively. Changes in (A) hepatic Ki-67+ cells, (B) hepatic TUNEL+ cells, (C) serum ALT level, (D) serum AST level, (E) serum TNF-α level, (F) serum IL-6 level, (G) hepatic EGF mRNA, and (H) hepatic OSM mRNA at day 7 of mice after indicated treatments. *p < 0.05, **p < 0.01, ***p < 0.001 mean significant changes between control and indicated treatment groups, respectively; #p < 0.05, ##p < 0.01, ###p < 0.001 mean significant changes between transplanted hADMSC treatment group and nontransplanted group, respectively; @p < 0.05, @@p < 0.01, @@@p < 0.001 mean significant change between nontreated group and treated with ZD or miR-210-KD treated with ZD, respectively. Ctrl, control; GL, Gal/LPS cotreatment; hADMSCs, human adipose-derived mesenchymal stem cells; KD, stable knockdown of miR-210; Gal, galactosamine; LPS, lipopolysaccharide; miR-210, microRNA-210; v-, vehicle-; Ki-67, antigen Ki-67; TUNEL, terminal deoxynucleotidyl transferase dUTP nick-end labeling; ALT, alanine transaminase; AST, aspartate transaminase; TNF, tumor necrosis factor; IL-6, interleukin-6; EGF, epidermal growth factor; OSM, oncostatin M.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Knockdown, Expressing, TUNEL Assay, Control, Derivative Assay

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: ZD ameliorates PKC/Raf-1/MAPK/NF-κB pathway activity, which was activated by LPS/H2O2 in hADMSCs. (A-C) PKC activity, NF-κB (p65), and phosphorylated Elk1 level were measured after LPS/H2O2 incubation with or without ZD cotreatment. (D) The expression level of miR-210 was measured after LPS/H2O2 incubation with or without ZD cotreatment. (E) Western blot results of Raf-1, phosphorylated MEK, total MEK, phosphorylated p38 MAPK, and total p38 MAPK after LPS/H2O2 incubation with or without ZD cotreatment. *p < 0.05, **p < 0.01, 096368916X694274p < 0.001 mean significant changes between control and treatment groups, respectively; #p < 0.05, ###p < 0.001 mean significant changes between indicated treatment and LPS/H2O2 incubation group, respectively. (E) Different letters (e.g., a vs. b, b vs. c) indicate a significant change between compared groups (p < 0.05). The same letters (e.g., a vs. a, c vs. c) indicate there is no significant change between compared groups (p > 0.05). Ctrl, control; ZD, zeaxanthin dipalmitate; L+H, LPS/H2O2; v-ZD, vehicle-ZD treatment; hADMSCs, human adipose-derived mesenchymal stem cells; LPS, lipopolysaccharide; PKC, protein kinase C; NF-κB, nuclear factor κB; Elk1, ETS domain-containing protein; miR-210, microRNA-210; Raf-1, RAF proto-oncogene serine/threonine protein kinase; MEK, mitogen-activated protein kinase; p38, p38 mitogen-activated protein kinase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Activity Assay, Incubation, Expressing, Western Blot, Control, Derivative Assay

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: ZD improves stress-induced cell damage and the level of miR-210 through the PKC/MAPK pathway in hADMSCs. (A–D) Cell viability, apoptotic ratio, secretion of TNF-α, and miR-210 expression were measured after LPS/H2O2 incubation with or without ZD cotreatment, MAPK inhibitor PD98059, or PKC inhibitor staurosporine. White column: control group, dark gray column: LPS/H2O2 coincubation group, light gray column: vehicle-PD98059 or vehicle-staurosporine group, gray column: LPS/H2O2 + ZD, PD98059, or staurosporine group. **p < 0.01, ***p < 0.001 mean significant changes between control and treatment groups, respectively; ##p < 0.01, ###p < 0.001 mean significant changes between indicated treatment and LPS/H2O2 incubation group, respectively; @p < 0.05, @@p < 0.01 mean significant changes between LPS/H2O2 + ZD group and LPS/H2O2 + ZD + PD98059 or LPS/H2O2 + ZD + staurosporine groups, respectively. ZD, zeaxanthin dipalmitate; hADMSCs, human adipose-derived mesenchymal stem cells; LPS, lipopolysaccharide; TNF-α, tumor necrosis factor-α; miR-210, microRNA-210.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Expressing, Incubation, Control, Derivative Assay

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: Cross-regulation between miR-210 and cellular ROS generation through ISCU1/2 in hADMSCs. (A–C) miR-210 mimic induced the generation of cellular and mitochondrial ROS, while ZD ameliorated the increase of those ROS productions. (D) After miR-210 mimic treatment, the production of ISCU was significantly decreased, while ZD recovered it. (E–H) When ISCU was knocked down by specific siRNA, both cellular and mitochondrial ROS were induced but inhibited by the ZD cotreatment. (C) and (G) Higher MitoSOX readout means more severe accumulation of mitochondrial ROS. ***p < 0.001 means significant change between indicated treatment groups. Ctrl, control; ZD, zeaxanthin dipalmitate; ROS, reactive oxygen species; miR-210, microRNA-210; mitoSOX, mitochondrial superoxide indicator; ISCU1/2, iron–sulfur cluster assembly proteins 1/2; siRNA, small interfering RNA; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Control, Small Interfering RNA

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: The expression level of miR-210 in hADMSCs is directly regulated by cellular ROS production and ZD treatment. (A, B) The expression of miR-210 was dose-dependently induced by ROS generator antimycin or rotenone after different incubation durations. (C) miR-210 expression was measured in hADMSCs when treated with antimycin, rotenone, LPS/H2O2 with or without NAC, a kind of ROS scavenger. (D) ZD cotreatment successfully reduced the elevated miR-210 expression by antimycin or rotenone. **p < 0.01, ***p < 0.001 mean significant changes between control and treatment groups, respectively; ##p < 0.01, ###p < 0.001 mean significant changes between added NAC and non-added NAC group or between nontreated and treated with ZD group, respectively. Ctrl, control; ZD, zeaxanthin dipalmitate; hADMSCs, human adipose-derived mesenchymal stem cells; LPS, lipopolysaccharide; NAC, N-acetyl-L-cysteine; miR-210, microRNA-210.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Expressing, Incubation, Control, Derivative Assay

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: Maintenance of normal miR-210 expression is critical for cell homeostasis of hADMSCs. (A) We constructed replication-deficient lentivirus-encoding human miR-210 precursor or inhibitor sponge to overexpress (OE) or knock down (KD) the endogenous expression of miR-210 in hADMSCs, respectively. (B, C) Cell viability and apoptotic ratio were measured after OE or KD of miR-210 in stem cell pretreated with LPS/H2O2 or not, in the absence or presence of ZD. (D) Treated OE or KD of miR-210 in stem cell with LPS/H2O2 with or without ZD altered the level of miR-210. Dark gray column: LPS/H2O2 coincubation, dark columns: overexpressed miR-210 with or without LPS/H2O2 treatment, gray columns: LPS/H2O2 + overexpressed miR-210 + ZD cotreatment, light gray columns: knocked down miR-210 with or without LPS/H2O2 treatment. *p < 0.05, **p < 0.001 mean significant changes between control and indicated treatment groups, respectively; #p < 0.05, ##p < 0.01, ###p < 0.001 mean significant changes between LPS/H2O2 treatment group and OE/KD miR-210 group, respectively; @p < 0.05, @@p < 0.01, @@@p < 0.001 mean significant change between non-treated and treated with ZD group, respectively. Ctrl, control; miR, microRNA; LPS, lipopolysaccharide; ZD, zeaxanthin dipalmitate; hADMSCs, human adipose-derived mesenchymal stem cells.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Expressing, Construct, Knockdown, Control, Derivative Assay

Journal: Cell Transplantation

Article Title: Precise Regulation of miR-210 is Critical for the Cellular Homeostasis Maintenance and Transplantation Efficacy Enhancement of Mesenchymal Stem Cells in Acute Liver Failure Therapy

doi: 10.3727/096368916X694274

Figure Lengend Snippet: ZD significantly increased and stable knockdown of miR-210 impaired the therapeutic efficacy of transplanted hADMSCs in a murine ALF model, respectively. (A) Mice survival numbers were recorded for 7 days with indicated treatments. (B, C) Representative hepatic H&E images (200x) and corresponding liver necrosis quantification of mice at day 1 and day 7 that received indicated treatments. White dashed line defined areas that are typical necrotic and inflammatory sites in the liver. (D) Expanded cell number in the host mice liver after stem cell transplantation. *p < 0.05, **p < 0.01, ***p < 0.001 mean significant changes between indicated groups, respectively. Ctrl, control; GL, Gal/LPS cotreatment; hADMSCs, human adipose-derived mesenchymal stem cells; KD, stable knockdown of miR-210; Gal, galactosamine; LPS, lipopolysaccharide; miR-210, microRNA-210; v-, vehicle-.

Article Snippet: Stem Cell Culture and Treatments hADMSCs were purchased from and validated by Cyagen Biosciences (Guangzhou, P.R.

Techniques: Knockdown, Drug discovery, Transplantation Assay, Control, Derivative Assay