Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Morphological characterization of bone marrow-derived mesenchymal stem cells. A: The morphological appearance of bone marrow-derived mesenchymal stem cells (BMSCs) at passage 4 of cell culture (magnification 40 ×); B-F: Expression of green fluorescence protein 72 h after BMSCs transfection by Ad-uPA at different levels of multiplicity of infection (MOI) (MOI = 20, 40, 60, 80, and 100; magnification 40 ×).

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Derivative Assay, Cell Culture, Expressing, Fluorescence, Transfection, Infection

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Western blot detection of urokinase plasminogen activator protein expression in bone marrow-derived mesenchymal stem cells after Ad- urokinase plasminogen activator transfection. Lane 1, uninfected bone marrow-derived mesenchymal stem cells (BMSCs); Lane 2, BMSCs infected with recombinant adenovirus containing human urokinase plasminogen activator (uPA) cDNA; Lane 3, BMSCs infected with control adenovirus. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Western Blot, Expressing, Derivative Assay, Transfection, Infection, Recombinant, Control

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Comparison of functional liver levels and liver fibrosis parameters in serum

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Comparison, Functional Assay, Control

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Histopathological change of liver tissue in different groups. A: HE and Masson staining was used to detect structural changes in liver tissues; B: Quantitative analyzes of liver fibrosis were performed using Masson stained sections. Five random views from each sample in each group were analyzed. BMSCs: Bone marrow-derived mesenchymal stem cells; uPA: Urokinase plasminogen activator.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Staining, Derivative Assay

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Effect of urokinase plasminogen activator gene modified-bone marrow-derived mesenchymal stem cells transplantation on the mRNA expression of molecules of the Wnt signaling pathway. The bar graph shows mean relative mRNA expression levels of β-catenin, Wnt4 and Wnt5a in liver tissues. Each sample was repeated three times from each cluster. Data are normalized to GAPDH mRNA expression levels. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; BMSCs: Bone marrow-derived mesenchymal stem cells; uPA: Urokinase plasminogen activator.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Modification, Derivative Assay, Transplantation Assay, Expressing

Journal: World Journal of Gastroenterology

Article Title: Human urokinase-type plasminogen activator gene-modified bone marrow-derived mesenchymal stem cells attenuate liver fibrosis in rats by down-regulating the Wnt signaling pathway

doi: 10.3748/wjg.v22.i6.2092

Figure Lengend Snippet: Effect of urokinase plasminogen activator gene modified-bone marrow-derived mesenchymal stem cells transplantation on the protein expression of molecules of the Wnt signaling pathway. A: Representative Western blot analysis of uPA, β-catenin, Wnt4, and Wnt5a protein expression levels in four groups; B: The bar graph represents mean relative protein expression levels of urokinase plasminogen activator, β-catenin, Wnt4, and Wnt5a. Data are normalized to GAPDH protein expression levels. GAPDH: Glyceraldehyde 3-phosphate dehydrogenase; BMSCs: Bone marrow-derived mesenchymal stem cells; uPA: Urokinase plasminogen activator.

Article Snippet: Cell culture and subculture BMSCs derived from the SD rat were purchased from Sciencell Research Laboratories (San Diego, California, United States).

Techniques: Modification, Derivative Assay, Transplantation Assay, Expressing, Western Blot

Journal: Scientific Reports

Article Title: SMAD3 and SMAD4 have a more dominant role than SMAD2 in TGFβ-induced chondrogenic differentiation of bone marrow-derived mesenchymal stem cells

doi: 10.1038/srep43164

Figure Lengend Snippet: Following lentiviral transduction with either SMAD2-shRNA, SMAD3-shRNA, SMAD4-shRNA or non-mammalian shRNA control (Ctrl-shRNA), human fetal BMSCs were cultured in chondrogenic medium for 1 day. Short hairpin RNA-mediated knockdown was evaluated by determining gene (RT-qPCR; ( a , c , e ) and protein (Western blot; ( b , d , f ) expression of SMAD2 ( a , b ), SMAD3 ( c , d ) and SMAD4 ( e , f ). Protein levels were normalized to GAPDH and expressed as relative to Ctrl-shRNA. Gene expression was normalized to the mean C T value of RPS27a and TBP . Data are expressed as % relative to normalized mRNA levels in Ctrl-shRNA. Bars represent mean + S.D. of triplicate pellets from 2 experiments. **p < 0.01; ***p < 0.001 compared to Ctrl-shRNA.

Article Snippet: Human fetal BMSCs (#SCC7500, Lot#6890, ScienCell Research Laboratories, Carlsbad, CA, USA) were expanded in Mesenchymal Stem Cell Growth Medium (MSCGMTM; Lonza, Basel, Switzerland) supplemented with 1% Penicillin-Streptomycin-Glutamine (Gibco, Carlsbad, CA, USA).

Techniques: Transduction, shRNA, Control, Cell Culture, Knockdown, Quantitative RT-PCR, Western Blot, Expressing, Gene Expression

Journal: Scientific Reports

Article Title: SMAD3 and SMAD4 have a more dominant role than SMAD2 in TGFβ-induced chondrogenic differentiation of bone marrow-derived mesenchymal stem cells

doi: 10.1038/srep43164

Figure Lengend Snippet: Human fetal BMSCs were transduced with adenovirus overexpressing SMAD2 (ad-SMAD2), ad-SMAD3, ad-SMAD4 or ad-LacZ as control, followed by culturing in chondrogenic medium for 1 day. To confirm overexpression, gene (RT-qPCR; ( a , c , e ) and protein (Western blot; ( b , d , f ) expression of SMAD2 ( a , b ), SMAD3 ( c , d ) and SMAD4 ( e , f ) were measured. Protein levels were normalized to GAPDH and expressed as relative to ad-LacZ. Gene expression data are presented as −ΔC T compared to the mean C T value of RPS27a and TBP . Bars represent mean + S.D. of triplicate pellets from 2 experiments. ***p < 0.001 compared to ad-LacZ.

Article Snippet: Human fetal BMSCs (#SCC7500, Lot#6890, ScienCell Research Laboratories, Carlsbad, CA, USA) were expanded in Mesenchymal Stem Cell Growth Medium (MSCGMTM; Lonza, Basel, Switzerland) supplemented with 1% Penicillin-Streptomycin-Glutamine (Gibco, Carlsbad, CA, USA).

Techniques: Transduction, Control, Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Gene Expression

Journal: Scientific Reports

Article Title: SMAD3 and SMAD4 have a more dominant role than SMAD2 in TGFβ-induced chondrogenic differentiation of bone marrow-derived mesenchymal stem cells

doi: 10.1038/srep43164

Figure Lengend Snippet: Human fetal BMSCs were transduced (lentivirus) either with SMAD2-shRNA, SMAD3-shRNA, SMAD4-shRNA or Ctrl-shRNA and subsequently pellet-cultured for 1, 7 or 14 days in chondrogenic medium with TGFβ. The effect of shRNA-mediated knockdown of SMAD2, SMAD3 and SMAD4 on chondrogenesis was determined by RT-qPCR analysis of the chondrogenesis-specific genes; ACAN ( a ), COL2A1 ( b ) and SOX9 ( c ), deposition of glycosaminoglycans (GAGs) per pellet ( d ) and per μg DNA ( e ), histological examination of GAGs by Safranin O (Saf.O) staining and collagen type II by immunohistochemistry in sections of 14 days-cultured pellets ( f ), and macroscopic evaluation of 14 days-cultured pellets ( g ). In ( f ) representative images of consecutive pellet sections per condition are shown and the scale bar represents 500 μm. Gene expression data are presented as −ΔC T compared to the mean C T value of RPS27a and TBP . Bars represent mean + S.D. of triplicate pellets from 2 experiments. *p < 0.05; **p < 0.01; ***p < 0.001 compared to Ctrl-shRNA.

Article Snippet: Human fetal BMSCs (#SCC7500, Lot#6890, ScienCell Research Laboratories, Carlsbad, CA, USA) were expanded in Mesenchymal Stem Cell Growth Medium (MSCGMTM; Lonza, Basel, Switzerland) supplemented with 1% Penicillin-Streptomycin-Glutamine (Gibco, Carlsbad, CA, USA).

Techniques: shRNA, Cell Culture, Knockdown, Quantitative RT-PCR, Staining, Immunohistochemistry, Gene Expression

Journal: Scientific Reports

Article Title: SMAD3 and SMAD4 have a more dominant role than SMAD2 in TGFβ-induced chondrogenic differentiation of bone marrow-derived mesenchymal stem cells

doi: 10.1038/srep43164

Figure Lengend Snippet: Human fetal BMSCs transduced either with adenoviral SMAD2 (ad-SMAD2), ad-SMAD3, ad-SMAD4 or ad-LacZ as control were pellet-cultured for 1, 7 or 14 days in chondrogenic medium with TGFβ. The effect of SMAD overexpression on chondrogenesis was determined by RT-qPCR analysis of the chondrogenesis-specific genes; ACAN ( a ), COL2A1 ( b ) and SOX9 ( c ), deposition of glycosaminoglycans (GAGs) per pellet ( d ) and per μg DNA ( e ), histological examination of GAGs by Safranin O (Saf.O) staining and collagen type II by immunohistochemistry in sections of 14 days-cultured pellets ( f ), and macroscopic evaluation of 14 days-cultured pellets ( g ). In ( f ) representative images of consecutive pellet sections per condition are shown and the scale bar represents 500 μm. Gene expression data are presented as −ΔC T compared to the mean C T value of RPS27a and TBP . Bars represent mean + S.D. of triplicate pellets from 2 experiments. *p < 0.05; **p < 0.01; ***p < 0.001 compared to ad-LacZ.

Article Snippet: Human fetal BMSCs (#SCC7500, Lot#6890, ScienCell Research Laboratories, Carlsbad, CA, USA) were expanded in Mesenchymal Stem Cell Growth Medium (MSCGMTM; Lonza, Basel, Switzerland) supplemented with 1% Penicillin-Streptomycin-Glutamine (Gibco, Carlsbad, CA, USA).

Techniques: Control, Cell Culture, Over Expression, Quantitative RT-PCR, Staining, Immunohistochemistry, Gene Expression