摘要
目的:探讨不同胚龄人神经干细胞分离和培养的最适条件,为神经干细胞的基础及应用研究打下基础。 方法:采用无血清培养和单细胞克隆技术,从胚龄10周和20周人脑海马组织中分离和培养神经干细胞,并应用免疫细胞化学染色予以鉴定。 结果:从两个不同胚龄的人脑海马组织中均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10周龄的人胚脑海马组织在高密度悬浮培养法时神经干细胞生长不良,在其他密度及培养法均有神经干细胞克隆球形成;20周的人胚脑海马组织仅在中等密度悬浮培养法及 高密度贴壁培养法中有克隆球形成;包被组两种胚龄海马组织在各种密度培养法神经干细胞均生长不良。 结论:人胚脑海马组织中存在具有自我更新和多分化潜能的神经干细胞,且胚龄越大,培养难度越大;高密度贴壁培养法适宜各胚龄神经干细胞的分离培养。
AIM: To investigate the suitable culture conditions of isolation culture and identification of human neural stem cells from different embryonic brains for further basic research and clinical application.
METHODS: Cells from human fetal hippocampus at embryonic age 10 week and 20 week were collected and cultured with serum free medium and single cell clone determination, then identified for immunocytochemical staining. RESULTS: The human neural stem cells having the abilities of self-renewal and multipotency were successfully isolated and cultured from two different human fetal hippocampuses. In the uncoated group, the cells from human fetal hippocampus at embryonic age 10-week could form clones in different ways excepting high dense suspension culture, and in 20 week brain, the cells could form clones on it in middle dense suspension culture and high dense adhere wall culture. In the coated group, the cells couldn't form clones. CONCLUSION: Neural stem cells exist in human fetal hippocampus. With fetal age increasing, then neural stem cells become more and more difficult to be isolated, high dense adhere wall culture can be used in embryonic brains with different fetal age.
出处
《中国临床康复》
CSCD
2004年第9期1726-1727,共2页
Chinese Journal of Clinical Rehabilitation
基金
广东省重点攻关项目(B30502)~~
