组织块法体外培养人胚胎神经干细胞

目的：探讨体外分离、培养人胚胎脑组织神经干细胞的实用方法。方法：从人胚胎脑额叶皮层分离组织块，采用DMEM／F12＋N2无血清培养基，进行体外培养、传代。采用免疫细胞化学法鉴定神经干细胞和分化后的其他细胞表型。结果：从人胚胎脑额叶皮层分离的细胞具有增殖和多向分化潜能，可进行传代培养，获得的细胞中大部分为Nestin表达阳性的细胞。诱导分化后可以出现NSE、GFAP表达阳性的细胞。结论：用上述方法分离培养的细胞具有神经干细胞的特性，并表达Nestin蛋白，认为是神经干细胞。此方法是一种具有实用价值的体外培养人胚胎脑神经干细胞的技术。

Tissue Piece Culture of Human Embryonic Neural Stem Cells in Vitro

Objective To explore a practiacl method of tissue piece culture of neural stem cells in brain frontal lobe of human embryo. Methods The serum free culturing technic was performed to culture and passage neural stem cells from embryonic human brain frontal lobe. Immunocytochemistry was used to identify the neural stem cells and their potentiality of differentiation. Results Nestin-positive neural stem cells and the neurospheres could be cultured and grown in the medium of DMEM/F12+N2 with EGF?bFGF. After attachment they can differentiate into GFAP and NSE positive cells. Conclusion The method to isolate and culture neural stem cells from embryonic human brain frontal lobe is available and valuable. The cultured cells are of features of neural stem cells and express Nestin so that may be considered as the neural stem cells.