GFP转基因胚胎大鼠神经干细胞生物学特性研究

目的 体外分离、培养、鉴定绿色荧光蛋白（green fluorescent protein，GFP）转基因胚胎大鼠神经干细胞，研究其体外活性与生物学性状，为在体分化示踪研究奠定基础。方法 采用机械分离，无血清传代培养法从GFP胚胎大鼠海马获得神经干细胞，免疫荧光和免疫细胞化学法染色进行体外神经干细胞性质和分化活性鉴定，并利用流式细胞计数法行纯度检测。结果 成功从GFP胎鼠海马获得神经干细胞，体外生长情况与活性良好，能分化成神经元和胶质细胞；体外连续传代3次后绝大部分细胞Nestin表达阳性，且分化后细胞的GFP表达不受影响。结论 GFP胚胎大鼠来源神经干细胞具有普通胎鼠NSCs相同的自我更新和多向分化潜能，且能稳定表达GFP，因此它可以成为示踪神经干细胞研究的有效工具细胞。

The Biological Characters of the Neural Stem Cells of Embryonic GFP Transgenic rat

Objetive: To stdudy the activity and biological characters of embryonic neural stem cells(NSCs) by isolating,incubating and identificating the NSCs from GFP transgenic rat in vitro, so as to establish a basis for the tracer study of differentiation of NSCs. Methods: NSCs were obtained from the hippocampus of the embryonic GFP transgenic rat and cultured in serum-free medium. The biological characters and activity were identified by immunochemistry and observed with fluoroscope.The purity of NSCs was detected by FACS. Results: The cells from embryonic hippocampus growing well,which were capable of self-renewal and differentiating into neurons and astrocytes.After the third passage most of the cells expressed Nestin. The expression of GFP protein did not change in neurospheres and differentiated cells. Conclusion: The NSCs harvested from embryonic GFP transgenic rat have the abilities of self-renewal, multipotency of differentiation and stable expression of GFP, so these cells may be valuable for the tracer study of NSCs.