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: Stem Cell Research & Therapy

Article Title: M2 macrophages promote heterotopic ossification through MSX2 binding to LEF1-mediated endothelial-mesenchymal transition

doi: 10.1186/s13287-026-04916-7

Figure Lengend Snippet: M2 macrophages promote osteogenic differentiation of MACECs via MSX2 regulation. A AR staining and B ALP staining of MAOECs cultured in osteogenic differentiation medium for 14 days. Original magnification: ×100. C Protein expression of OPN, OCN, and Runx2 of MAOECs cultured in osteogenic differentiation medium for 14 days detected by western blotting. D mRNA expression of OPN, OCN, and Runx2 of endothelial cells cultured in osteogenic differentiation medium for 14 days detected by qPCR. E Alcian Blue staining of MAOECs cultured in chondrogenic differentiation medium for 14 days. Scale bar: 100 μm. F Protein expression of SP7 and SOX9 of MAOECs cultured in chondrogenic differentiation medium for 14 days detected by western blotting. G mRNA expression of SP7 and SOX9 of MAOECs cultured in chondrogenic differentiation medium for 14 days detected by qPCR. **P < 0.01, ***P < 0.001. All in vitro experiments were repeated three times. Full-length blots are presented in Figure 3 of Original Images for Blots file

Article Snippet: For osteogenic/chondrogenic differentiation, after the above grouping, the MAOECs were further cultured in osteogenic differentiation induction medium (HS-B-001, Hasen, Wuxi, China)/chondrogenic differentiation induction medium (PD-005, Procell, Wuhan, China) with added cytokines.

Techniques: Staining, Cell Culture, Expressing, Western Blot, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Extracellular Vesicles Derived from Human Umbilical Cord Mesenchymal Stem Cells Alleviated the Inflammatory Response in Mice Infected with the Influenza Virus A (H1N1)

doi: 10.3390/ijms26188839

Figure Lengend Snippet: Characterization of human umbilical cord mesenchymal stem cells (hUCMSCs) and extracellular vesicles derived from hUCMSCs (hUCMSC-EVs). ( A ) hUCMSCs passed to the 3rd generation grew adherently to the wall and were in good condition, with the morphology of elongated pike/pike shape and the overall distribution of vortex (Scale bar = 200 μm). ( B ) Oil red O staining of hUCMSCs after lipogenic induction, visible with obvious red lipid droplets (Scale bar = 100 μm). ( C ) Alizarin red staining of hUCMSCs after osteogenic induction, visible with obvious red calcium nodules (Scale bar = 100 μm). ( D ) Identification of hUCMSCs surface markers by flow cytometry. ( E ) ZETAVIEW particle potential titration and particle size analyzer for hUCMSC-EVs particle size detection. ( F ) Transmission electron microscopy to observe hUCMSC-EVs morphology (scale bar = 200 nm), the red arrow indicates the hUCMSC-EVs. ( G ) Western blotting to identify the expression of hUCMSC-EVs marker protein.

Article Snippet: When the cell density reached 80–90%, the cells were cultured in adipogenic differentiation induction medium (Dakewe, Beijing, China, #6114531) and osteogenic differentiation induction medium (Dakewe, Beijing, China, #6114541) for 18–21 days.

Techniques: Derivative Assay, Staining, Flow Cytometry, Titration, Transmission Assay, Electron Microscopy, Western Blot, Expressing, Marker