摘要
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神经干细胞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。
Objective To study the activity and biological characteristics of the embryonic neural stem cells （NSCs） from green fluorescent protein （GFP） transgenic rat by in vitro isolation, incubation, and identification of these cells and to lay the foundation for the research on tracking differentiation. Methods NSCs from the hippocampus of the embryonic GFP transgenic rat were generated by mechanical isolation and serum-free culture. The biological characteristics and differentiated activity were identified by immunochemistry and observed under a fluoroscope. The purity of NSCs was detected by FACS. Results The cells from the hippocampus of the transgenic rat were successflally generated and they grew well, had ideal activity, and were capable of differentiating into neurons and neuroglia. By the third passage, most of the cultured cells expressed Nestin and the expression of GFP protein did not change in the neurospheres and differentiated cells. Conclusions The NSCs harvested from embryonic GFP transgenic rat have the similar ability to self-renew and the similar multipotency in differentiation as those from the ordinary embryonic rat and they stably express GFP. The cells may be used as an effective tool for the research on tracking NSCs.
出处
《实用医学杂志》
CAS
2008年第5期709-712,共4页
The Journal of Practical Medicine
基金
上海市教委自然科学基金资助项目(编号:04BB23)
关键词
绿色荧光蛋白质类转
基因
胚胎
神经干细胞
生物学特性
Green fluorescent proteins Transgenic Embryonic Neural stem cell Biological characteristics
