Journal: Stem Cell Reviews and Reports

Article Title: Bone Marrow Mesenchymal Stromal Cell Osteogenesis is driven by Paracrine signals from Regulatory T Cell

doi: 10.1007/s12015-025-11015-2

Figure Lengend Snippet: Optimization of culture conditions. a Viable and non-viable regulatory T cells (Treg) from one donor cultured in RPMI or TexMACS media, with and without starvation, at days 14 and 21. b Characterization of Treg cultured in TexMACS or RPMI with and without starvation, at days 14 and 21. c Phalloidin staining for F-actin (red), DAPI staining for nuclei (blue), and STRO-1 expression (green). Scale bar: 50 μm. d Metabolic activity of bone marrow mesenchymal stromal cells (BMSC) cultured in growth media (GM) or TexMACS. Data are presented as mean ± standard error of mean from three independent donors ( n = 3) * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001

Article Snippet: Cells were fixed with 4% paraformaldehyde (Sigma-Aldrich) for 10 min, permeabilized (Invitrogen) for 30 min, and blocked with 3% BSA and 10% Normal Goat Serum (Thermo Fisher Scientific) in DPBS for 1 h. Cells were then incubated overnight at 4 °C with anti-STRO-1 antibody (1:150) (Santa Cruz Biotechnology, USA) in 3% BSA/DPBS, and followed by Alexa FluorTM 488 secondary antibody (1:200) (Thermo Fisher Scientific) in DPBS for 1 h. Nuclei and cytoskeleton was stained with DAPI (1:1000) (Thermo Fisher Scientific) and Phalloidin Atto 565 (1:500) (Sigma-Aldrich) in DPBS for 1 h. Images were taken by Dragonfly 505 confocal microscope (Andor Technologies Inc, Northern Ireland).

Techniques: Cell Culture, Staining, Expressing, Activity Assay

Journal: Stem Cell Reviews and Reports

Article Title: Bone Marrow Mesenchymal Stromal Cell Osteogenesis is driven by Paracrine signals from Regulatory T Cell

doi: 10.1007/s12015-025-11015-2

Figure Lengend Snippet: Effect of Treg condition medium (Treg-CM; 50 µg/mL) or non-condition medium (Non-CM; 50 µg/mL) on bone marrow mesenchymal stromal cells (BMSC) metabolic activity, morphology, and migration. a BMSC metabolic activity on days 1, 4, and 7. b BMSC treated with Treg-CM on day 4, with GM and Non-CM as controls. Phalloidin staining for F-actin (red), DAPI staining for nuclei (blue), and STRO-1 expression (green). Scale bar: 50 μm. c Migration of BMSC treated with Treg-CM. d Quantification of BMSC migration e Crystal violet staining of migrated BMSC treated with Treg-CM after 72 h, with GM and Non-CM as controls. Data are presented as mean ± standard error of mean from three independent donors ( n = 3). * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001

Article Snippet: Cells were fixed with 4% paraformaldehyde (Sigma-Aldrich) for 10 min, permeabilized (Invitrogen) for 30 min, and blocked with 3% BSA and 10% Normal Goat Serum (Thermo Fisher Scientific) in DPBS for 1 h. Cells were then incubated overnight at 4 °C with anti-STRO-1 antibody (1:150) (Santa Cruz Biotechnology, USA) in 3% BSA/DPBS, and followed by Alexa FluorTM 488 secondary antibody (1:200) (Thermo Fisher Scientific) in DPBS for 1 h. Nuclei and cytoskeleton was stained with DAPI (1:1000) (Thermo Fisher Scientific) and Phalloidin Atto 565 (1:500) (Sigma-Aldrich) in DPBS for 1 h. Images were taken by Dragonfly 505 confocal microscope (Andor Technologies Inc, Northern Ireland).

Techniques: Activity Assay, Migration, Staining, Expressing