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: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: Co-culture with SSCs rescues the function of irradiated osteogenic precursor cells. (A, B) Cell apoptosis was analyzed by flow cytometry with Annexin V-PE/7AAD double staining. (A) Representative flow cytometry plots. (B) Quantitative analysis of the apoptotic rate. (C, D) ALP activity was assessed. (C) Representative ALP staining images (Scale bar: 50 μm). (D) Quantitative analysis of the relative ALP activity. (E, F) Mineralization capacity was evaluated using Alizarin Red S staining. (E) Representative staining images of mineralized nodules (Scale bar: 100 μm). (F) Quantitative analysis of the relative mineralization level. (G, H) Cell migration was determined by a migration assay. (G) Representative images of migrated cells (Scale bar: 50 μm). (H) Quantitative analysis of the relative cell migration level. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01).

Article Snippet: After irradiation and corresponding interventions, cells were cultured in osteogenic induction medium (iXCells Biotechnologies, San Diego, CA, Cat. No. MD-0006) for 7 days.

Techniques: Co-Culture Assay, Irradiation, Flow Cytometry, Double Staining, Activity Assay, Staining, Migration, Two Tailed Test

Journal: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: SSCs exert rescue effects via the Wnt/β-catenin signaling pathway. (A) Representative ALP staining images of cells in each group (Scale bar: 50 μm). (B) Quantitative analysis of ALP activity in each group. (C) Representative Alizarin Red S staining images of cells in each group (Scale bar: 100 μm). (D) Quantitative analysis of Alizarin Red S staining in each group. (E) Relative mRNA expression levels of osteogenic marker genes ( Runx2 , Col1a1 , and OCN ) detected by qRT-PCR. GAPDH was used as an internal reference gene. (F) Representative Western blot images showing the expression levels of RUNX2, COL1A1, OCN, and β-catenin in each group. GAPDH was used as a loading control. (G) Quantitative analysis of Western blot results (gray value ratio of target protein to GAPDH) in each group. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: After irradiation and corresponding interventions, cells were cultured in osteogenic induction medium (iXCells Biotechnologies, San Diego, CA, Cat. No. MD-0006) for 7 days.

Techniques: Staining, Activity Assay, Expressing, Marker, Quantitative RT-PCR, Western Blot, Control, Two Tailed Test

Journal: Dose-Response

Article Title: Skeletal Stem Cells Rescue Radiation-Induced Osteogenic Precursor Cell Dysfunction via the Wnt/β-Catenin Signaling Pathway

doi: 10.1177/15593258261440983

Figure Lengend Snippet: SSCs alleviate the radiation-induced bone injury in mice. (A–G) Micro-CT analysis of bone microstructure. (A) Representative micro-CT images of femurs. Quantitative analysis of (B) bone mineral density (BMD), (C) bone volume fraction (BV/TV), (D) trabecular thickness (Tb.Th), (E) trabecular number (Tb.N), (F) connectivity density (Conn.D), and (G) trabecular separation (Tb.Sp) at 2- and 4-weeks post irradiation. (H–K) Histological analysis (Scale bar: 100 μm). (H) H&E staining showing steatosis (arrows) and (I) quantitative analysis of steatotic lesions per field. (J) TRAP staining showing osteoclasts (arrows) and (K) quantitative analysis of osteoclast number per field. (L–O) Immunohistochemical staining of osteogenic markers (Scale bar: 100 μm). (L) Osterix staining and (M) quantitative analysis of Osterix-positive area. (N) β-catenin staining and (O) quantitative analysis of β-catenin-positive area. All experiments were conducted in three groups: Control, irradiation (IR), and IR plus SSC (IR+SSC) at 2- and 4-weeks post-irradiation. All data are presented as mean ± SD, with statistical significance determined by unpaired two-tailed Student’s t-test (* p < 0.05; ** p < 0.01; *** p < 0.001)

Article Snippet: After irradiation and corresponding interventions, cells were cultured in osteogenic induction medium (iXCells Biotechnologies, San Diego, CA, Cat. No. MD-0006) for 7 days.

Techniques: Micro-CT, Irradiation, Staining, Immunohistochemical staining, Control, Two Tailed Test

Journal: Molecular medicine reports

Article Title: MicroRNA‑125b suppresses the proliferation and osteogenic differentiation of human bone marrow‑derived mesenchymal stem cells.

doi: 10.3892/mmr.2014.2024

Figure Lengend Snippet: Figure 1. Comparison of the proliferation and osteogenic differen tiation of hBMSCs isolated from osteoporotic patients and normal subjects. (A) Growth and viability of hBMSCs were determined by the MTT assay after cells were cultured for 2, 4, 6, 8 and 10 days; (B) hBMSCs at passage 3 were cultured in osteogenic medium for 14 days, followed by staining and assessment of ALP activity. Data were analyzed using the Student's t‑tests; (C) quantitative polymerase chain reaction analysis of miR‑125b expression in hBMSCs. miR‑125b expression levels were increased in hBMSCs isolated from elderly patients. Data were subjected to Student's t‑tests. Error bars represent the mean ± standard deviation of three independent experiments. *P<0.05. hBMSCs, human bone marrow‑derived mesenchymal stem cells; ALP, alkaline phosphatase; miR, micro RNA; OD, optical density.

Article Snippet: On every third day, the medium was replaced with osteogenic differentiation medium (10% fetal bovine serum, Hyclone; 100 nM dexamethasone, 45 mM L-ascorbic acid and 10 mM β-glycerophosphate; Sigma, St. Louis, MO, USA).

Techniques: Comparison, Isolation, MTT Assay, Cell Culture, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Expressing, Standard Deviation

Journal: Molecular medicine reports

Article Title: MicroRNA‑125b suppresses the proliferation and osteogenic differentiation of human bone marrow‑derived mesenchymal stem cells.

doi: 10.3892/mmr.2014.2024

Figure Lengend Snippet: Figure 3. (A) Overexpression of miR‑125b suppressed the osteogenic differentiation of hBMSCs. Osteoblast differentiation of hBMSCs was induced by osteogenic differentiation medium and hBMSCs were collected 14 days after osteogenic induction. qPCR analysis measured the expression of Runx‑2, ALP, OC and COL1α1 in hBMSCs. qPCR data were subjected to Student's t‑tests. Error bars represent the mean ± standard deviation of three independent experi ments. *P<0.05. (B) ALP staining was performed after 14 days of culture to detect ALP activity and Alizarin Red staining was performed after 21 days to evaluate mineralized bone matrix formation. (C) ALP activity of hBMSCs was determined 14 days after osteogenic induction by p‑Nitrophenyl Phosphate Liquid Substrate System and absorbance was measured at 405 nm. Data were subjected to Student's t‑test. *P<0.05. hBMSCs, human bone marrow‑derived mesenchymal stem cells; qPCR, quantitative polymerase chain reaction; Runx‑2, Runt‑related transcription factor‑2; ALP, alkaline phosphatase; OC, osteo calcin; COL1α1, collagen type I α1; NC, negative control; miR, micro RNA; OD, optical density.

Article Snippet: On every third day, the medium was replaced with osteogenic differentiation medium (10% fetal bovine serum, Hyclone; 100 nM dexamethasone, 45 mM L-ascorbic acid and 10 mM β-glycerophosphate; Sigma, St. Louis, MO, USA).

Techniques: Over Expression, Expressing, Standard Deviation, Staining, Activity Assay, Real-time Polymerase Chain Reaction, Negative Control

Journal: Heliyon

Article Title: Hepatocellular carcinoma patients serum modulates the regenerative capacities of adipose mesenchymal stromal cells

doi: 10.1016/j.heliyon.2024.e24794

Figure Lengend Snippet: Effect of HCC serum treatment for 6 days on the adipogenic, osteogenic, and angiogenic potential of hA-MSCs. (A, B) The adipogenic differentiation potential was observed in normal serum-treated hA-MSCs (A) compared to HCC serum (B). Arrow indicates the formation of Oil red O-stained oil droplets in response to adipogenic induction. (C,D) The osteogenic differentiation potential was observed in normal serum treated-hA-MSCs (C) compared to HCC serum (D). Arrow indicates the formation of orange-stained Ca 2+ deposits in response to osteogenic induction. (E – H) CAM assay for evaluating the angiogenesis in-ovo 5 days post-implantation in (E) Normal serum-treated hA-MSCs, (F) HCC serum-treated hA-MSCs. (G – H) Isolated CAM from the egg with (G) Normal serum-treated hA-MSCs, (H) HCC serum-treated hA-MSCs. (I) Normalized gene expression analysis using real-time PCR in normal and HCC serum-treated hA-MSCs. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To induce the osteogenic differentiation, hA-MSCs were cultured in an osteogenic differentiation medium (DMEM, 10 % FBS, 2 mM l -glutamine, 100 U/ml penicillin-streptomycin amphotericin, 0.1 μM dexamethasone, 50 μM l -ascorbic acid, and 10 mM β-glycerophosphate) for 21 days (Lonza, USA; Serva, Germany; Sigma Aldrich, USA).

Techniques: Staining, Chick Chorioallantoic Membrane Assay, In Ovo, Isolation, Gene Expression, Real-time Polymerase Chain Reaction

Journal: Journal of Cellular Biochemistry

Article Title: Oxidative-Mechanical Stress Signals Stem Cell Niche Mediated Lrp5 Osteogenesis in eNOS −/− null mice

doi: 10.1002/jcb.24031

Figure Lengend Snippet: Panel A. The myofibroblast cells stain for Alcian blue, indicating a cartilaginous phenotype treated with osteogenic media.

Article Snippet: Mineralization Assay Porcine valvular myofibroblast cells were grown for extended periods (42 days) in Osteogenic Media (Stem Cell Technologies, Vancouver, CA) to induce morphological changes with subsequent formation of bone-like nodules.

Techniques: Staining