人胚脊髓神经干细胞的分离培养和鉴定

The isolation, cultivation and identification of spinal cord-derived neural stem cells from human embryos

目的:探讨人胚脊髓神经干细胞的体外培养和分化的方法,观察其增殖和分化特点。方法:利用无血清培养和单细胞克隆技术从人胚脊髓组织中分离培养出神经干细胞并用血清诱导其分化,应用免疫荧光细胞化学技术对培养细胞及其分化细胞进行鉴定。结果:从人胚脊髓组织分离的细胞在EGF单独存在时无法形成神经球,在bFGF单独存在时只形成少量神经球,在EGF和bFGF共同存在时形成大量具有连续增殖能力的神经球,表达神经干细胞的标志物Nestin,经血清诱导后分化为神经元、星形胶质细胞和少突胶质细胞并表达特异性抗原NSE、GFAP和CNP。结论:在体外培养条件下可从人胚脊髓组织中培养出神经干细胞,它可为神经干细胞的基础研究和临床应用提供材料。

Objective: To explore the cultivation and differentiation in vitro of spinal cord-derived neural stem cells from human embryoes. Methods: Neural stem cells were isolated from the spinal cord of human embryoes with serum-free incubation and single cell cloning technique and were induced to differentiation by serum. Results: In the culture of human embryoes spinal cord,EGF alone does not produce any neurospheres. bFGF alone produces a few neurospheres. Both EGF and bFGF produce a lot of neurospheres with the potential to expand, and the cells expressed Nestin which were the markers of neural stem cells. Induced by serum, the neurospheres differentiated into neurons, astrocytes and oligodendrocytes and expressed NSE, GFAP and CNP,respectively. Conclusions: Neural stem cells could be cultured from spinal cord of human embryoes, and they could be applied in basic and clinic research.