Multilineage Differentiation of Murine MSCs

Murine bone marrow mesenchymal cells (10⁵ cells/well, 24-well plate) were incubated in an osteogenic medium (A1007201; Thermo Fisher Scientific Inc., Waltham, MA, USA) for 18 days. Mineralized extracellular matrices were probed using von Kossa staining [26 (link)]. For adipocyte formation, 10⁵ cells were incubated in adipogenic medium (A1007001; Thermo Fisher Scientific Inc., Waltham, MA, USA) for 15 days [22 (link)]. Adipocytes were probed using Nile red staining kit. In some experiments, a total of 10⁵ cells/well (24-well plates) were incubated in a differentiation medium for brown adipocytes (BADTM, Cosmo Bio Co., Ltd., Tokyo, Japan) and white adipocytes (WATDM, Cosmo Bio Co., Ltd., Tokyo, Japan) for 15 days, respectively, according to the maker’s instructions.