人胚神经干细胞的长期贴壁培养和鉴定

目的：探讨人胚神经干细胞的长期贴壁培养条件及传代方法.方法：在多种培养条件下,分别从9周和16周人胚脑中分离培养神经干细胞,应用免疫细胞化学的方法对所分离细胞的巢蛋白表达、自我更新能力和多向分化潜能进行鉴定.结果：采用高密度悬浮培养法能有效地从两种胚龄的人胚脑或室管膜下区中分离出神经干细胞,而采用贴壁培养法的人胚神经干细胞能保持干细胞的特性稳定增殖4个月.结论：从大小胚龄的人胚中均可成功分离培养人胚神经干细胞,贴壁培养的人胚神经干细胞能稳定增殖,是进一步进行人胚神经干细胞转基因研究的良好模型.

Long-term adhere-wall culture and identification of human embryonic neural stem cell

Objective:To establish the method of long-term adhere-wall culture and passage of human embryonic neural stem cell.Method:By means of different culture methods, neural stem cells were isolated from human miscarriaged embryonic brain (embryonic age 9 weeks and 16 weeks). The capabilities of self-renew?nestin expression and multipotentiality of the neural stem cells were isolated and identified with immunocytochemical staining.Result:Human embryonic neural stem cell with the capabilities of self-renewing and multipotentiality can be isolated from the brain or subependyma of human embryo at different embryonic age by high concentration suspension culture. Neural stem cells possessed their characters and proliferated for 4 months stably by adhere-wall culture.Conclusion:Human embryonic neural stem cell can be isolated from human embryo of different embryonic age successfully. Human embryonic neural stem cells cultured by adhere-wall method can proliferate stably for a long time, which serve as a good model for transgenic research of human embryonic neural stem cell.