人胚胎神经干细胞的分离培养及鉴定

Isolation and characterization of human embryos neural stem cell

目的研究人胚胎神经干细胞的培养条件及体外分化情况。方法将新鲜分离的胚胎（自然流产胎儿4-6周,经志愿者知情同意）无菌条件下取海马,采用无血清培养基进行培养,原代培养的细胞增殖成神经干细胞球,倒置显微镜观察细胞的形态学,并用免疫荧光方法对神经干细胞进行自我更新能力（Brd U）的鉴定及特异性抗原Nestin的检测。结果培养5-7d的神经干细胞球呈桑葚样悬浮生长,呈明显的Brd U阳性反应,其具有增殖能力;神经干细胞表达了Nestin抗原阳性。结论人胚胎神经干细胞具有神经干细胞的基本特征,可进一步用于基础及临床研究。

Objective To study the culture condition and in vitro differentiation of human neural stem cells. Methods The cells isolated from fresh human fetal hippocampus（4-6 weeks） were seeded into defined serum-free medium and cultured, Inverted microscope was used to observe the cell morphology, and immunofluorescence method was used to observe self-renewal ability of the cells labeled with Brd U and the specificity of Nestin antigen of the neural stem cells. Results The harvested cells grew as neurosphere in mulberry suspension and passed serially, and presented Brd U positively and specific antigen Nestin positively. Conclusion The human embryonic neural stem cells have the basic characteristics of neural stem cells and can be used for basic and clinical research further.