All differentiation assays were performed during passage 2. To promote osteogenic and adipogenic differentiation, 1x105 were cultured in StemXVivo™ Human/Mouse Osteogenic/Adipogenic Base Media (R&D, Minneapolis, USA) supplemented with penicillin-streptomycin; after 21 days, the plates were fixed with 4% paraformaldehyde and stained by Alizarin Red and Sudan Black, respectively. For chondrogenic differentiation, 3x105 cells were placed in Falcon tubes with 5 mL of StemXVivoTM Human/Mouse Chondrogenic (R&D). After 3 days, the cells were resuspended in 2 mL of differentiation medium (R&D). After 21 days, the pellets were fixed, stained using Alcian Blue and Masson Trichrome, and analyzed.9 (link)
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