Journal: PLoS ONE

Article Title: Lung Fibroblasts Share Mesenchymal Stem Cell Features Which Are Altered in Chronic Obstructive Pulmonary Disease via the Overactivation of the Hedgehog Signaling Pathway

doi: 10.1371/journal.pone.0121579

Figure Lengend Snippet: (a)Rate of lung fibroblast conversion into myofibroblasts after 3, 4 and 5 days exposure to TGF-β1. * p<0.05 compared to untreated cells. # p<0.05 compared to non- smoker (C-NS) and smoker controls (C-S) exposed to TGF-β1 for 4 or 5 days. (b) Oil-red O staining of, MSC, DF and lung fibroblasts from C-NS, C-S and COPD subjects12 days after exposure to adipogenic differentiation medium. (c)Alcian blue staining of MSC, DF and lung fibroblasts at day 24 following chondrogenesis initiation. (d,e) (D, E) Comparative real-time PCR analysis of mRNA expression of genes encoding for (d) adipogenic peroxysome proliferative activated receptor gamma 2 (PPAR-γ 2; early), lipoprotein lipase (LPL; late) and (e)chondrogenic Sox-9 marker. (f) Alizarin red staining of cells following 17 days of osteoblastic induction and (g)real-time PCR analysis of mRNA expression of osteoblastic alkaline phosphatase (ALP, early) and osteocalcin (OC, late) markers. (d,e,g) Results represent means ± SEM of at least 3 independent experiments performed with donor fibroblasts. NS: non significant; * p<0.05, ** p<0.01. MSC, mesenchymal stem cells; DF, dermal fibroblast; C-NS, non-smokers controls; C-S, smoker controls; COPD, smokers with COPD.

Article Snippet: Chondrogenic differentiation was carried out using Mesenchymal Stem Cell Chondrogenic Differentiation Medium (PromoCell, Heidelberg, Germany).

Techniques: Staining, Real-time Polymerase Chain Reaction, Expressing, Marker

Journal: International Journal of Molecular Sciences

Article Title: A Comparative Study of Non-Viral Gene Delivery Techniques to Human Adipose-Derived Mesenchymal Stem Cell

doi: 10.3390/ijms150915044

Figure Lengend Snippet: Differentiation potential of hAD-MSCs and fibroblasts following transfection using four independent techniques. Osteogenic, adipogenic, and chondrogenic differentiation of ( A ) hAD-MSCs and ( B ) fibroblasts post-transfection using the microporation, standard electroporation, cationic polymer, and classical calcium phosphate precipitation transfection techniques. After 21 days, osteogenic cultures were stained using alizarin red. Adipogenic cultures were analyzed for lipid accumulation after 14 days of differentiation using inverted microscopy. Thereafter they were fixed and stained for triglycerides with Oil-Red-O. After 21 days, chondrogenic cultures were stained for spheroid formation using Alcian blue. (Magnification: ( A ) adipogenic 400×, osteogenic 40× and chondrogenic 40×; ( B ) adipogenic 400×, osteogenic 100× and chondrogenic 100×).

Article Snippet: The cells were induced to differentiate into chondrogenic cells by replacing the growth medium with chondrogenic differentiation medium (Promo Cell).

Techniques: Transfection, Electroporation, Staining, Inverted Microscopy

Journal: Molecular Medicine

Article Title: Aspirin attenuates the detrimental effects of TNF-α on BMMSC stemness by modulating the YAP-SMAD7 axis

doi: 10.1186/s10020-024-00890-z

Figure Lengend Snippet: Effect of TNF-α and aspirin on the stemness of BMMSCs.( A ) Representative pictures and quantitative results of colony formation of BMMSCs with different treatments; ( B , C ) The expression of stemness markers (C-MYC, NANOG, SOX2) of BMMSCs with different treatments were analyzed using qRT-PCR and western blot assays. ( D , E ) The expression of osteogenic differentiation markers (OCN, OPN, RUNX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( F , G ) The expression of chondrogenic differentiation markers (COL2A1, ACAN, SOX9) in BMMSCs from different groups were analyzed using qRT-PCR and western blot assays. Data in A-F are given as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 5 mm

Article Snippet: The cells were then inoculated into gelatin-coated 6-well plates and left to adhere for 24 h. Mesenchymal stem cell chondrogenic differentiation medium (ScienCell, USA) was then added to induce chondrogenic differentiation for 14 days, with medium changes every 3 days.

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Molecular Medicine

Article Title: Aspirin attenuates the detrimental effects of TNF-α on BMMSC stemness by modulating the YAP-SMAD7 axis

doi: 10.1186/s10020-024-00890-z

Figure Lengend Snippet: Inhibition of YAP reverses the protective effect of AS on BMMSC stemness ( A , B ) YAP in different treatment groups was determined using qRT-PCR and western blot assays with or without the AS and TNF-α treatments; ( C , D ) Detection of YAP knockdown efficiency using qRT-PCR and western blot assays; ( E ) Representative pictures and quantitative results of colony formation of BMMSCs with different treatments; ( F , G ) The expression of stemness markers (C-MYC, NANOG, SOX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( H , I ) The expression of osteogenic differentiation markers (OCN, OPN, RUNX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( J , K ) The expression of chondrogenic differentiation markers (COL2A1, ACAN, SOX9) in BMMSCs with different treatments were analyzed using qRT-PCR and western blot assays. Data in A-K are given as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 5 mm

Article Snippet: The cells were then inoculated into gelatin-coated 6-well plates and left to adhere for 24 h. Mesenchymal stem cell chondrogenic differentiation medium (ScienCell, USA) was then added to induce chondrogenic differentiation for 14 days, with medium changes every 3 days.

Techniques: Inhibition, Quantitative RT-PCR, Western Blot, Knockdown, Expressing

Journal: Molecular Medicine

Article Title: Aspirin attenuates the detrimental effects of TNF-α on BMMSC stemness by modulating the YAP-SMAD7 axis

doi: 10.1186/s10020-024-00890-z

Figure Lengend Snippet: Overexpression of YAP attenuates the damaging effects of TNF-α on BMMSC stemness ( A , B ) Detection of YAP overexpression efficiency using qRT-PCR and western blot assays; ( C ) Representative pictures and quantitative results of colony formation of BMMSCs with different treatments; ( D , E ) The expression of stemness markers (C-MYC, NANOG, SOX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( F , G ) The expression of osteogenic differentiation markers (OCN, OPN, RUNX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( H , I ) The expression of chondrogenic differentiation markers (COL2A1, ACAN, SOX9) in BMMSCs from different groups were analyzed using qRT-PCR and western blot assays. Data in A-I are given as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 5 mm

Article Snippet: The cells were then inoculated into gelatin-coated 6-well plates and left to adhere for 24 h. Mesenchymal stem cell chondrogenic differentiation medium (ScienCell, USA) was then added to induce chondrogenic differentiation for 14 days, with medium changes every 3 days.

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

Journal: Molecular Medicine

Article Title: Aspirin attenuates the detrimental effects of TNF-α on BMMSC stemness by modulating the YAP-SMAD7 axis

doi: 10.1186/s10020-024-00890-z

Figure Lengend Snippet: Overexpression of SMAD7 attenuates the protective effects of YAP on the stemness of BMMSCs ( A ) The prediction of proteins that may bind to YAP was carried out using the STRING website; ( B ) Co-IP experiments were conducted to detect YAP binding to SMAD7; ( C , D ) qRT-PCR and western blot experiments were performed to detect SMAD7 expression in BMMSCs after overexpression of YAP; ( E , F )Detection of SMAD7 overexpression efficiency using qRT-PCR and western blot assays G Representative pictures of colony formation of BMMSCs with different treatments; ( H , I ) The expression of stemness markers (C-MYC, NANOG, SOX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( J , K ) The expression of osteogenic differentiation markers (OCN, OPN, RUNX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( L , M ) The expression of chondrogenic differentiation markers (COL2A1, ACAN, SOX9) in BMMSCs with different treatments were analyzed using qRT-PCR and western blot assays. Data in C-M are given as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 5 mm

Article Snippet: The cells were then inoculated into gelatin-coated 6-well plates and left to adhere for 24 h. Mesenchymal stem cell chondrogenic differentiation medium (ScienCell, USA) was then added to induce chondrogenic differentiation for 14 days, with medium changes every 3 days.

Techniques: Over Expression, Co-Immunoprecipitation Assay, Binding Assay, Quantitative RT-PCR, Western Blot, Expressing

Journal: Molecular Medicine

Article Title: Aspirin attenuates the detrimental effects of TNF-α on BMMSC stemness by modulating the YAP-SMAD7 axis

doi: 10.1186/s10020-024-00890-z

Figure Lengend Snippet: Inhibition of SMAD7 reverses the damaging effects of TNF-α on BMMSC stemness ( A , B ) Detection of SMAD7 knockdown efficiency using qRT-PCR and western blot assays; ( C )Representative pictures and quantitative results of colony formation of BMMSCs with different treatments; ( D , E ) The expression of stemness markers (C-MYC, NANOG, SOX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( F , G ) The expression of osteogenic differentiation markers (OCN, OPN, RUNX2) of BMMSCs in different groups were analyzed using qRT-PCR and western blot experiments. ( H , I ) The expression of chondrogenic differentiation markers (COL2A1, ACAN, SOX9) in BMMSCs from different groups were analyzed using qRT-PCR and western blot assays. Data in A-I are given as the mean ± SD of three independent experiments. * P < 0.05, ** P < 0.01, *** P < 0.001. Scale bars: 5 mm

Article Snippet: The cells were then inoculated into gelatin-coated 6-well plates and left to adhere for 24 h. Mesenchymal stem cell chondrogenic differentiation medium (ScienCell, USA) was then added to induce chondrogenic differentiation for 14 days, with medium changes every 3 days.

Techniques: Inhibition, Knockdown, Quantitative RT-PCR, Western Blot, Expressing