摘要
目的:旨在研究hTERT基因所表达的蛋白对NSCs生物学特性的影响。方法:通过脂质体转染法将构建的重组质粒pEGFP-N1-hTERT转染到大鼠胎儿NSCs中,对转染的NSCs进行体外诱导分化试验、裸鼠致瘤性试验、RT-PCR、Western blot和流式细胞仪分析。结果:转染的NSCs在体外培养中仍具有干细胞的生长特性和多潜能性、无致瘤性,转染重组质粒的NSCs中,存在hTERT基因mRNA的转录产物和融合蛋白hTERT-GFP的表达。结论hTERT基因对神经干细胞端粒酶活性有上调作用,可促进NSCs的体外增殖和永生化趋势。
Objective： researching on the influence biological characteristics. Method：Constructed recombinant of protein expressed by hTERT gene on the NSCs plasmid pEGFP-NI-hTERT was transfected into rat fetal NSCs with lipid-mediated transfection method-Transfected NSCs differentiation in vitro tests, tumorigenicity in nude mice test, RT-PCR and Western-blot analysis and flow cytometry were carried on. Result Transfected NSCs stem cells still have the potential growth characteristics and muhipotency in vitro, no tumorigenic, and transcription product of hTERT gene mRNA and hTERT-GFP fusion protein expression in the transfected NSCs. Conclusions： The transferred NSCs of telomerase activity was up-regulation and the proliferation of cells were promoted in vitro by hTERT gene, and also displayed immortalization tendency.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2008年第6期7-12,共6页
China Biotechnology
基金
国家“863”计划资助项目(2002AA216161)
