Journal: Stem Cells International

Article Title: Proteomic Analysis Reveals Commonly Secreted Proteins of Mesenchymal Stem Cells Derived from Bone Marrow, Adipose Tissue, and Synovial Membrane to Show Potential for Cartilage Regeneration in Knee Osteoarthritis

doi: 10.1155/2021/6694299

Figure Lengend Snippet: Representative characterization of bone marrow (BM), adipose tissue (AT), and synovial membrane (SM) mesenchymal stem cells (MSCs). (a) Phase-contrast microscopy images of MSCs (scale bar = 100 μ m). (b) Differentiation potential of MSCs. Adipogenic differentiation (Oil Red O, top), osteogenic differentiation (Alizarin Red S, middle), and chondrogenic differentiation (Alcian Blue, bottom). (c) The immunophenotyping of MSCs using flow cytometry. Positive (CD73, CD90, and CD105) and negative (CD14, CD34, CD45, CD79a, and human leukocyte antigen-DR (HLA-DR)) markers. The histogram is shown with an overlay isotype control.

Article Snippet: After 2 days, differentiation was induced using chondrogenic differentiation medium (containing Dulbecco's minimal EM-high glucose (HyClone), 10% FBS, 1% P/S, 0.1 μ M of dexamethasone, 1X insulin-transferrin-selenium (Gibco), 50 μ M of L-ascorbic acid 2-phosphate sesquimagnesium salt hydrate, and 5 ng/mL of transforming growth factor β 1 (TGF- β 1; PeproTech, Rocky Hill, NJ, USA)) [ ].

Techniques: Microscopy, Flow Cytometry

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: The Journal of Veterinary Medical Science

Article Title: Isolation and characterization of equine dental pulp stem cells derived from Thoroughbred wolf teeth

doi: 10.1292/jvms.16-0131

Figure Lengend Snippet: Representative images of in vitro differentiated eDPCs. Alizarin red staining of control (a) and osteogenic induction (b) cultures. Alcian blue staining of control (c) and chondrogenic induction (d) cultures. Scale Bar, 50 µ m (a–d). Oil red O staining of control (e) and adipogenic induction (f) cultures. Scale Bar, 5 µ m (e, f).

Article Snippet: For chondrogenic differentiation, passage 3 eDPCs were seeded in 6-well culture plates at a density of 2.5 × 10 3 cells/cm 2 , and after incubation in PM for 24 hr, the medium was changed to chondrogenic induction medium (Differentiation Basal Medium-Chondrogenic, Lonza) supplemented with 4.5 g/ l D -glucose, 350 μ M L -proline, 100 nM dexamethasone and 0.02 g/ l transforming growth factor (TGF)-β3.

Techniques: In Vitro, Staining, Control

Journal: Stem Cell Reports

Article Title: Equine-Induced Pluripotent Stem Cells Retain Lineage Commitment Toward Myogenic and Chondrogenic Fates

doi: 10.1016/j.stemcr.2015.12.005

Figure Lengend Snippet: Equine iPSC Intrinsic Propensities to Muscle and Cartilage Derivatives in Teratomas (A) Representative pictures of derivatives (arrowheads) of ectoderm (upper, ectodermal cysts; lower, neural tube), endoderm (unstructured, gland-like acinar tissues), and mesoderm (upper, adipose tissue; lower, immature bone structures). (B) Quantitation of striated muscle patches (white arrows) by means of Masson trichrome staining ( ∗ p < 0.05, Mann-Whitney U test, n = 3 independent experiments per cell type). (C) Quantitation of chondrogenic patches (black arrows) by means of Alcian blue staining ( ∗ p < 0.05, Mann-Whitney U test, n = 3 independent experiments per cell type). (D) Immunofluorescence staining for lamin A/C on equine iPSC teratomas (n = 4/cell type; ct-, negative control, murine iPSC teratoma; ct+, positive control, human iPSC teratoma). (E) Staining for lamin A/C and markers of ectodermal (TUJ1), endodermal (αFP), and mesodermal (SARCαACT) derivatives in equine MAB-iPSC-derived teratomas (n = 4 technical replicates per cell type). Magnifications of highlighted fields are shown to the right of original panels to evidence co-localization of lamin A/C (nuclear membrane) and the cytoplasmic markers. Comparable results were obtained with MSC-iPSCs (data not shown). Histograms represent average values; error bars indicate SD. Scale bars, 100 μm.

Article Snippet: To initiate chondrogenic differentiation, 3 × 10 5 cells were resuspended in 5 ml of medium and centrifuged for 5 min at 150 × g . Subsequently, 0.5 ml of chondrogenic differentiation medium (basal differentiation medium [Lonza] supplemented with 10 ng/ml TGF-β 3 [Sigma]) or expansion medium were added per pellet and refreshed twice a week.

Techniques: Quantitation Assay, Staining, MANN-WHITNEY, Immunofluorescence, Negative Control, Positive Control, Derivative Assay, Membrane

Journal: Stem Cell Reports

Article Title: Equine-Induced Pluripotent Stem Cells Retain Lineage Commitment Toward Myogenic and Chondrogenic Fates

doi: 10.1016/j.stemcr.2015.12.005

Figure Lengend Snippet: Assessment of Equine iPSC Chondrogenic Differentiation (A) Alcian blue staining for undifferentiated and chondrogenic microspheres. Indicated percentages refer to the Alcian blue-positive areas (arrows) across the depicted spheres. (B) Quantitation of cell density and differentiation efficiency in parental cells and iPSCs, comparing undifferentiated (undiff) and differentiated (chondro) spheres ( ∗ p < 0.05, one-way ANOVA with Bonferroni comparison; ns, not significant). (C) qRT-PCR with equine-specific COMP primers; data are depicted as fold change versus undiff (AU, arbitrary units; nd, not detectable; ∗ p < 0.05, one-way ANOVA with Bonferroni comparison; n = 3 independent experiments [3 technical replicates per experiment/cell type]). (D) Oil Red staining on MAB- and MSC-iPSCs after adipogenic differentiation (red lipid vacuoles denote adipocyte-like cells). Differentiation rate is quantitated as percentage of adipocytes per field ( ∗ p < 0.05, Mann-Whitney U test, n = 3/cell type, n = 3 independent experiments per cell type). Histograms represent average values; error bars indicate SD. Scale bars, 100 μm.

Article Snippet: To initiate chondrogenic differentiation, 3 × 10 5 cells were resuspended in 5 ml of medium and centrifuged for 5 min at 150 × g . Subsequently, 0.5 ml of chondrogenic differentiation medium (basal differentiation medium [Lonza] supplemented with 10 ng/ml TGF-β 3 [Sigma]) or expansion medium were added per pellet and refreshed twice a week.

Techniques: Staining, Quantitation Assay, Comparison, Quantitative RT-PCR, MANN-WHITNEY