Journal: Frontiers in Endocrinology

Article Title: Steroidogenic differentiation of human amniotic membrane-derived mesenchymal stem cells into a progesterone-/androgen-producing cell lineage by SF-1 and an estrogen-producing cell lineage by WT1−KTS

doi: 10.3389/fendo.2024.1410433

Figure Lengend Snippet: Differentiation capacity of hAmMSCs. (A) Adipogenic differentiation was evidenced by Oil Red O staining. Lipid vacuoles were formed in the hAmMSCs. (B) Osteogenic differentiation was evidenced by Alizarin Red S staining. Mineralized matrix was formed in the hAmMSCs. (C) The hAmMSCs cultured with chondrogenic differentiation medium formed pellets. Note that the cells did not form pellets in the control medium (RPMI 1640 plus 10% Fetal Calf Serum). Chondrogenic differentiation was evidenced by Alcian blue staining. Scale bar, 100 µm. hAmMSCs, human amniotic membrane-derived mesenchymal stem cells.

Article Snippet: To induce osteogenic differentiation, the amniotic membrane-derived cells were seeded in Mesenchymal Stem Cell Osteogenic Differentiation Medium (PromoCell) at a density of 2×10 4 cells/cm 2 and cultured for 1 week, replacing the medium every 3 days.

Techniques: Staining, Cell Culture, Control, Membrane, Derivative Assay

Journal: Scientific reports

Article Title: Analysis of cell-biomaterial interaction through cellular bridge formation in the interface between hGMSCs and CaP bioceramics.

doi: 10.1038/s41598-020-73428-y

Figure Lengend Snippet: Figure 6. Osteoinductible effect of the CaP bioceramic on hGMSCs in culture. (A,B,C,D) Representative fluorescent Confocal microscopy image showing a CaP bioceramic seeded with hGMSCs after 5 h in culture (T0) in the absence of osteogenic induction. In blue, Hoechst nuclear staining (A), in green actin (B) and red OPN (C). Merge of 3 channels is shown in (D). Images represent maximum projections from Z-Stack reconstructions obtained with Confocal Microscopy. Scale bar is 200 μm. (E,F,G,H) Representative fluorescent Confocal microscopy image showing a CaP bioceramic seeded with GMSCs after 1 week in culture (T1) in the absence of osteogenic induction. In blue, Hoechst nuclear staining (E), in green actin (F) and red OPN (G). Merge of 3 channels is shown in (H). Images represent maximum projections from Z-Stack reconstructions obtained with Confocal Microscopy. Scale bar is 200 μm. (I,J,K,L) Representative fluorescent Confocal microscopy image showing a CaP bioceramic seeded with hGMSCs showing an increase in OPN expression after 4 weeks in culture (T2) in the absence of osteogenic induction. In blue, Hoechst nuclear staining (I), in green actin (J) and red OPN (K). Merge of 3 channels is shown in (L). Images represent maximum projections from Z-Stack reconstructions obtained with Confocal Microscopy. Note the dense cellular network at this time, with cells spreading outside the CaP bioceramic (L, stars). Scale bar is 200 μm. (M) Graph depicts relative expression of OPN in hGMSCs seeded with CaP bioceramics at day 21 obtained by qPCR, showing an increase in mRNA expression in OPN as compared to the control condition, differences were statistically significant *P < 0.05 verified by Mann Whitney U test, Non-parametric test N = 3 biological replicates with two technical replicates each.

Article Snippet: After indicated incubation periods hGMSCs on CaP bioceramics were fixed with 4% para-formaldehyde/PBS and subjected to immunofluorescence analysis for ki67 ab-66155 (Santa Cruz) (expression cell proliferation) or for OPN ab-8448 (Abcam) (osteogenic cell differentiation) and actin sc-8432 (Santa Cruz) for cell cytoskeleton.

Techniques: Confocal Microscopy, Staining, Expressing, Control, MANN-WHITNEY

Journal: Stem Cell Reports

Article Title: WNT3A Promotes Hematopoietic or Mesenchymal Differentiation from hESCs Depending on the Time of Exposure

doi: 10.1016/j.stemcr.2013.04.002

Figure Lengend Snippet: WNT3A-Induced Mesospheres Differentiate to Osteogenic, Adipogenic, and Smooth Muscle Lineages (A) Heatmap showing that WNT3A- and BIO-induced mesospheres were enriched for transcripts associated with chondrocyte and bone differentiation. The expression of most of these genes differed in sorted d4 WNT3A/BMP GFP + and GFP − fractions. The scale in arbitrary units is shown. (B–G) Immunocytochemical and histochemical analysis of WNT3A-induced mesodermal colonies recultured and subjected to mesenchymal stem cell differentiation toward osteogenic and adipogenic fates. FABP-4 (B) and oil red O (G) reactivity confirmed adipogenic potential and osteogenic potential was confirmed by expression of OSTEOCALCIN (C, arrows) and alizarin red staining (F). The presence of smooth muscle was shown by SMA immunoreactivity (D). (E) IgG control for (B)–(D). Scale bars, 50 μM (B and D), 100 μM (C and G), and 200 μM (E and F). (H) Model illustrating that the effects of WNT3A during mesoderm differentiation depend on the time of exposure. WNT3A synergizes with BMP4 to increase mesodermal differentiation. Methylcellulose cultures from BMP4- and WNT3A/BMP4-induced EBs stimulated with VEGF without addition of WNT3A or BIO form hemangioblast colonies, while cultures with WNT3A or BIO do not form hematopoietic colonies, but form mesospheres capable of differentiation toward bone, fat, and smooth muscle. See also .

Article Snippet: Colonies formed in MC cultures supplemented with 100 ng/ml WNT3A or 5 μM (2′Z,3′E)-6-bromoindirubin-3′-oxime (BIO) (Calbiochem) were assayed for osteogenic and adipogenic potential using the Human Mesenchymal Stem Cell Functional Identification Kit (R&D Systems) according to the manufacturer’s instructions.

Techniques: Expressing, Cell Differentiation, Staining, Control