Isolation, Culture, Adipogenisis And Osteogenesis Differentiation Of Rabbit Bone Marrow Mesenchymal Stem Cells In Vitro

Objective: To investigate the isolation and cultivation of bone marrow mesenchymal stem cells < BMSCs ) from the bone marrow of rabbit and induce them to differentiate into adipocytes and osteoblasts in vitro. Methods: BMSCs were harvested from the femurs, tibias and humerus bones of the rabbit, then separated, purified and proliferated. After primary culture, the subcultured cells were cultured in adipogenisis inducing medium including dexamethasone, insulin, isobutylxanthine and 3-isobutyl-l-methyl-xanthine, then stained with Oil Red O. The subcultured cells were also cultured in osteogenesis inducing medium including dexamethasone, |3-glycerophosphate, L-ascorbic acid, then stained with NBT/BCIP and alizarin red. Results: The BMSCs of rabbit showed active proliferative capability in primary and passage cultures. After 72 hours of adipocyte inducing, the BMSCs displayed accumulation of lipid vacuoles. On the 21th day, approximately 80% BMSCs were induced to adipocytes as detected by Oil Red O. After 8 days of osteoblast inducing, the BMSCs congregated , showed typical cuboidal shape and formed mineralized nodule. The expression of alkaline phosphatase was positive. Conclusion: The BMSCs from rabbit bone marrow showed stable, long-term growth in vitro, rapid proliferation, easily anchorage and could be passaged continuously in present culture condition. The BMSCs can be induced to adipogenisis and osteogenesis committed differentiation. It indicates that the BMSCs could be exploited as the seeding cells in tissue engineering and be used for the stem cells pharmacology.