成年脊髓损伤大鼠脊髓神经干细胞的体外培养及分化研究

目的 探讨大鼠脊髓损伤后脊髓神经干细胞的分离培养方法及分化情况.方法 采用Allen法制作大鼠脊髓损伤模型,利用无血清培养和单细胞克隆技术在成年脊髓损伤7 d大鼠脊髓中分离具有单细胞克隆能力的神经干细胞,并进行培养鉴定.结果 从成年脊髓损伤7 d大鼠脊髓中成功分离出神经干细胞,该细胞具有连续克隆能力,可传代培养,表达神经巢蛋白抗原.分化后的细胞表达神经元细胞、星形胶质细胞和少突胶质细胞的特异性抗原.结论 致伤7 d的成年大鼠脊髓组织体外町培养出神经十细胞,并分化为神经无细胞、星形胶质细胞和少突胶质细胞,有可能参与脊髓损伤的修复过程.

Culture and differentiation of neural stem cells from the injured spinal cord in adult rats

Objective To investigate culture and differentiation of neural stem cells from the injured spinal cord in adult rats.Methods The spinal cord of male SD rats was injured by Allen's weight dropping on T8.Procedures of floating neural stem cell culture were carried out.Results Neurospheres of neural stem cells emerged after plating for 1 week.They had potent serf-renewing ability and could be passaged repeatedly,Immunohistochemistry data showed that these neurospheres strongly expressed nestin.A characteristic intermediate filament was seen in neural stem cells.The stem cells were further induced to differentiate into cells expressing β-tubulin,glial fibrillary acidic protein (GFAP) and myelin associated glycopretein (MAG),suggesting their differentiating potencies into neurons,astrocytes and oligodendrocytes.Conclusion In adult rats after spinal cord injury the neural stem cells can be isolated and cultured in vitro and induced to differentiate into neurons,astrocytes and oligodendrocytes,so the neural stem cells may participate in repair of spinal cord injury.