体外培养获取高纯度神经前体细胞

Gaining high purified neural precursor cells in culture

目的 获取较高纯度的神经前体细胞 (NPC)。方法 加入 30ng/mlbFGF培养E12天大鼠皮质、海马和隔区的NPC ,用免疫组化与免疫荧光组化方法将其显示。结果 加bFGF组的NPC不断分裂 ,形成大量的分裂球并维持到第 4代 ;绝大部分细胞 (约 98%以上 )表达Nestin ,约有 6 0 %为BrdU和Nestin免疫荧光双标阳性细胞。对照组的分裂球维持时间较短 ,随培养时间延长细胞向外迁移生长 ;第 2代约有 6 0 %细胞表达Nestin ,40 %为BrdU和Nestin免疫荧光双标阳性细胞。结论 bFGF对E12天大鼠脑组织的NPC有明显的促分裂作用 ,选用胎龄较小的胚脑组织加入适量的bFGF培养可获取较高纯度的NPC。

Objective To gain high purified neural precursor cells in culture.Methods The neural precursor cells(NPCs)from embryonic day 12 of rats were cultured in the present of bFGF(30ng/ml)and detected by immunoperoxidase and immunoflueonce staining.Results The NPC,formed a lot of spheres of proliferation.sustained to proliferate in the present of bFGF till 4 passages.The majority of the cells,about more than 98%,express Nestin and 60% of them were positive cells with double immunoflueonce staining for anti BrDU and Nestin.The proliferation of the NPCs in the control group appeared for a short time and the cells of spheres of proliferation migrated outward with prolonging time in culture.About 60% of cells express Nestin and 40% of them were positive cells with double immunoflueonce staining for anti BrDU and Nestin.Conclusion BFGF obviously promoted the proliferation of NPCs of E12 rats.The high purified NPCs can be gained by culturing the brain cells of younger embryonic rats in the present of bFGF.