Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are able to differentiate into different cell lineages such as adipocytes and osteoblastes. Establishing a stable and reliable induction method to induce BMSCs to differentiate into adipocytes is of great help in studying prevention and treatment of osteoporosis and skeletal-related diseases.
OBJECTIVE: To investigate the optimal conditions for induced differentiation of BMSCs into adipocytes.
METHODS: Under aseptic condition, rat femur was harvested. BMSCs were isolated and purified by density-gradient centrifugation and adherent culture in vitro. After complete confluency, passage 3 BMSCs were induced to differentiate into adipocytes using DMEM containing 3-isobutyl-1-methylxanthine, indomethacin, dexamethasone, insulin and fetal bovine serum.
RESULTS AND CONCLUSION: Under the inverted microscope, highly homogenous BMSCs could be harvested after three passages, and these cells could be induced to differentiate into adipocytes. BMSCs could be committedly to differentiate into adipocytes in vitro after induced by 3-isobutyl-1-methylxanthine, indomethacin, dexamethasone. 5×10^-4 mol/L 3-isobutyl-1-methylxanthine, 2×10^-4 mol/L indomethacin and 10^-4 mol/L dexamethasone exhibited better effects on induced differentiation of BMSCs into adipocytes. The best condition for inducing BMSCs to differentiate into adipocytes in vitro was DMEM supplemented with 0.1 mmoL/L 3-isobutyl-1-methylxanthine, 10 mg/L insulin, 0.1 mmoL/L indomethacin, 1 μmoL/L dexamethasone, and 10% fetal bovine serum.