不同胚龄人神经干细胞的分离和培养

Isolation, culture and identification of human neural stem cells from different embryonic brains

目的 探索不同胚龄入神经干细胞分离和培养的适宜条件。方法 在多种培养条件下 ,采用无血清培养和单细胞克隆技术 ,从 7周和 13周人胚脑中分离和培养神经干细胞 ,应用免疫荧光染色予以鉴定。结果 从两个不同胚龄的人胚脑中分离出具有自我更新和多分化潜能的神经干细胞 ,未包被组 7周的人胚脑除高密度悬浮培养法生长不良 ,其它密度及培养法均有克隆球形成 ;13周的人胚脑仅中等密度悬浮培养法及高密度贴壁培养法有克隆球形成 ;包被组各种密度培养均生长不良。结论 人胚脑中存在具有自我更新能力和多分化潜能的神经干细胞 ,随着胚龄增大 ,培养难度逐渐加大 ,高密度贴壁培养法适宜各胚龄神经干细胞的分离培养。

Objective To isolate, culture and identify human neural stem cells from 7-week to 13-week embryonic brains and to study different suitable culture conditions.Methods By using serum-free cell culture and single cell clone test,the neural stem cells were isolated and cultured from 7-week human embryonic forebrains and 13-week human embryonic cerebral cortex in different conditions and immunofluorescence tests were used to identify the cells. Results The human neural stem cells having the abilities of self-renewal and multipotency were successfully isolated and cultured from different embryonic brains. In 7-week brain, the cells could form clones in different ways except in high concentration suspension culture.In 13-week brain, the cells could form clones only in middle concentration suspension culture and high concentration adhere-wall culture. In coated petri dish, the cells couldn't form clones.Conclusions There are neural stem cells in human embryonic brains. With gestational age increasing, the neural stem cells become more and more difficult to be isolated. High concentration adhere-wall culture can be used in embryonic brains with different gestation age.