大鼠骨髓间充质干细胞分离培养及向神经元样细胞分化的实验研究

Experimental Study on Isolation, Cultivation and Induced Differentiation of Rat Bone Marrow Stromal Stem Cells

目的:探讨大鼠骨髓间充质干细胞(marrowstromalstemcell,MSC)的分离培养和体外扩增方法及生物学特性,研究其在诱导因子诱导下向神经元细胞分化的能力。方法:取大鼠骨髓,采用贴壁培养筛选法分离MSC,进行培养扩增,观察其生物学特性;用β-巯基乙醇(β-mereaptoethanol,β-ME)和丹参注射液诱导MSC向神经元样细胞分化,并通过细胞免疫化学法鉴定分化细胞。结果:大鼠MSC可通过贴壁法成功分离并可在体外大量扩增,经丹参注射液和β-ME诱导,MSC可向神经元样细胞分化,出现胞体和突起,细胞免疫化学染色神经元特异性烯醇化酶(NSE)和巢蛋白(Nestin)呈阳性。结论:MSC易分离和培养,体外培养条件下生长良好,可连续传代,诱导剂作用下可分化为神经元样细胞。

Objective： To investigate the method for the isolation and cultivation of rat bone marrow stromal stem cells （MSC） and the biological character of MSC, and study the possibility of inducing their differentiation into neuronlike ceils in vitro. Method： MSC was isolated from adult rats by wall sticking method. The proliferative characteristics were observed in primary and passage cultures. After differentiation induced by β-ME and danshen injection, eytoehemistry was used to identify the induced cells. Results： Rats MSC could be separated and amplified in vitro by wall sticking method. After being induced by β-ME and danshen injection for 5 hours, most of the cells would be differentiated into neuron-like cells, appearing cell body and apophysis. By eytoehemistry, cells showed positive of NSE and nestin. Conclusion： Rats MSC can grow well in vitro and be passaged continuously. MSC can be induced to differentiate to neuron-like cells.