成人骨髓间充质干细胞诱导分化为神经元样细胞后的维持培养

目的:探讨体外诱导成人骨髓间充质干细胞(bonemarrowstromalcells,BMSCs)分化为神经元样细胞后长期培养的可行性。方法:自成人红骨髓分离培养BMSCs,通过克隆培养、成骨分化、成脂肪分化鉴定其特性,取第3代细胞重新接种,经碱性成纤维生长因子预诱导及β-巯基乙醇诱导后,换维持培养液维持培养,观察其分化及生长情况。结果:成神经元诱导分化0.5h后,多数细胞即开始呈现出神经元样外观,5h后绝大多数细胞表现出典型的神经元样外观,维持培养1周后,大多数细胞继续维持神经元样外观,免疫组织化学染色示65%±3.3%的细胞微管相关蛋白-2染色阳性,43%±2.1%的细胞神经微丝-160染色阳性。结论:成人BMSCs经诱导后半数左右的细胞可分化为神经元样细胞,在优化的培养体系中,分化的神经元样细胞可存活1周以上。

Maintaining culture of neuron like cells inducted from human bone marrow stromal cells (BMSCs)

Objective:To find a protocol of sustaining culture of differentiated neuron like cells induced from human bone marrow stromal cells(BMSCs).Method:BMSCs were harvested from the marrow of adult volunteers and maintained in DMEM supplemented with 10% Fetal Bovine Serum,the character of BMSCs was identified by the generation of clone lines,osteogenic and lipogenic induction,the protocol of induced neuron like cells was established and the status of their maintaining culture was tested.Result:After half of an hour following neuronal induction some cells showed neuron like appearance,and 5 hours later almost all cells showed typical neuronal appearance.After 7 days,about half of the differentiated cells expressed the neuronal markers including microtubule-associated protein 2(MAP-2) and neurofilament protein(NF-160).Conclusion:With this induced culture protocol,BMSCs can differentiate into neuron like cells and keep alive for at least one week.