Stemxvivo human mouse osteogenic adipogenic base media

To promote osteogenic and adipogenic differentiation, 1 x 10 5 cells were cultured in StemXVivo™ Human/Mouse Osteogenic/Adipogenic Base Media (R&D, Minneapolis, USA) supplemented with penicillin-streptomyciun. When the osteogenic and adipogenic cells had reached approximately 70 and 100% confluence, respectively, differentiation medium was added (R&D, Minneapolis, USA). After 21 days, the cells were fixed with 4% paraformaldehyde and stained by Alizarin red and Sudan Black, respectively. For chondrogenic differentiation, 2.5 x 10 5 cells were placed in Falcon tubes with 5 mL StemXVivoTM Human/Mouse Chondrogenic medium (R&D) supplemented with penicillin-streptomyciun. After 3 days, the cells were resuspended in 2 mL differentiation medium (R&D). The pellets were maintained in medium containing a differentiator for 21 days and were then fixed, stained, and analyzed using Alcian Blue and Masson's trichrome staining. For neurogenic differentiation, 3 x 10 5 cells were placed on microscope slides. After reaching 80% confluence, 0.5 µL monoetilglicerol (Sigma) was added to each well. After 24 h, cells were fixed for immunocytochemical analysis.