脊髓源神经干细胞向胆碱能神经元诱导分化的研究

目的 研究胚胎大鼠脊髓源性神经干细胞的培养和在体外骨髓基质细胞诱导其分化的情况。方法 从孕龄13d的胚胎大鼠脊髓组织中分离获得神经干细胞，采用含表皮生长因子（EGF）及碱性成纤维细胞生长因子（bFGF）的无血清限定性培养基培养，同时进行诱导分化，观测脊髓神经干细胞向胆碱能神经元分化的情况，用细胞免疫荧光化学方法鉴定分化结果。结果 可从胚胎脊髓中分离得到大量的神经于细胞，通过限定性培养基培养可获得于细胞球，经从骨髓基质细胞培养液中获得的限制性培养液在体外可被诱导分化，用细胞免疫荧光化学法鉴定，可见有胆碱能神经元生成。结论 胚胎大鼠脊髓神经干细胞在添加EGF与bFGF的限定性培养基中可以增殖并长期保持稳定的性状，在体外可以被诱导分化为胆碱能神经元。

The study on the cholinergic differentiation of fetal spinal cord stem cells

Objective To study the cultivation of fetal spinal cord stem cells(SCSCs) in vitro,and its specific differentiation induced by bone marrow stromal cells(BMSCs).Methods The neural stem cells from spinal cord of 13 d fetal rats were harvested and cultivated in a serum-free limited medium which contained EGF and bFGF,and then were induced by conditioned medium obtained from the cultured medium of BMSCs.The results were identified by cellular immunohistochemistry.Results Lots of neural stem cells could be obtained from the fetal spinal cord,and the neurospheres were formed through cultivation in limited medium.The SCSCs could be induced to differentiate into cholinergic neurons by the cultured medium of BMSCs.Conclusion The limited medium can increase the population of SCSCs and maintain its attributes in vitro for a prolonged period with a stable genetic background.SCSCs can be induced to differentiate into cholinergic neurons.