兔骨髓间充质干细胞定向诱导分化为脂肪细胞的研究

Study on Rabbit Bone Marrow Mesenchymal Stem Cells Differentiated into the Adipocytes in Vitro

目的:探讨兔骨髓间充质干细胞(bonemesenchymalstemcells,BMSCs)体外分离培养、诱导分化为脂肪细胞的方法,并研究其生物学特性的变化.方法:采用体外细胞培养技术自兔股骨、胫骨及肱骨中分离BMSCs进行纯化、培养.用地塞米松、3-异丁基-1-甲基黄嘌呤(IBMX)、胰岛素、吲哚美辛定向诱导BMSCs分化为脂肪细胞,改良MTT法测定其生长曲线,油红O染色,光镜下观察橙红色脂滴沉着的细胞比例.结果:成脂诱导72h后,细胞内有脂滴出现,随着诱导时间的延长,脂滴逐渐增加并融合为脂泡;细胞由梭形转变为类圆形或多角形;第3周油红O染色显示80%以上的细胞转变为脂肪细胞.结论:兔BMSCs经体外分离、诱导培养可以定向分化为脂肪细胞.

Objective: To investigate the isolation and cultivation of bone mesenchymal stem cells (BMSCs) from the bone marrow of rabbit in vitro, and induce them to differentiate into adipocytes 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, 3-isobutyl-1-methyl-xanthine, insulin and isobutylxanthine, then stained with Oil Red O. Results: After 72 hours of adipocyte inducing, the BMSCs displayed accumulation of lipid vacuoles. On the 21th day, approximately 80% BMSCs were induced into adipogenic cells as detected by Oil Red O. Conclusion: The BMSCs isolated and cultured can be induced into adipogenisis committed differentiation.