Stemxvivo mouse rat osteogenic supplement 20

The induction of adipogenic differentiation was performed by culturing the cells in a homemade adipogenic differentiation medium for 21 days as already described (Doucet et al., 2005 (link)). Thereafter, we stained the cells using Oil Red for a duration of 5 min. For osteocyte differentiation, we purchased a specific medium containing P/S at 1:100 dilution + StemXVIVO Mouse/Rat Osteogenic supplement 20 × (R&D Systems). WT and TNFR2 KO-MSCs were cultured for 17 days. Then, they were stained for 3 min using 2% Alizarin red. Samples were visualized by the objectives of 4 × and 10 × of the microscope (EVOS XL Core, Invitrogen by Thermo Fisher Scientific).

WT and TNFR2 KO-MSCs were tested for their ability to differentiate into adipocytes and osteoblasts. Adipogenic differentiation was induced by culturing cells in a specific differentiation homemade medium for 21 days as already described [48 ]. Cells were then stained with Oil Red for 5 min. For osteocyte differentiation, MSCs were cultured in differentiation medium (StemXVIVO Osteogenic/Adipogenic Base Media supplemented with P/S/N at 1:100 dilution + StemXVIVO Mouse/Rat Osteogenic Supplement 20×) (R&D Systems) for 17 days and stained 3 min with 2% Alizarin Red.