外源性人端粒酶逆转录酶基因转染对正常人毛乳头细胞的影响

Effect of human telomerase reverse transcriptase gene transfection on human dermal papilla cells

目的研究外源性人端粒酶逆转录酶(human telomerase reverse transcriptase,hTERT)基因转染对体外培养正常人毛乳头细胞的影响。方法采用脂质体转染法,将hTERT基因导入体外培养的正常人毛乳头细胞,经G418筛选得到阳性克隆,连续传代培养。检测转染细胞内hTERT基因的整合和表达情况,初步分析转染细胞的生物学特性。结果转染后获得1个阳性细胞克隆,扩大培养后细胞生长良好,传代次数增加,已连续传至35代。检测证实转染细胞内hTERT基因在DNA、mRNA和蛋白质水平均有表达,且此转染细胞具有体外培养正常毛乳头细胞的生物学特性。结论外源性hTERT基因转染可以延长体外培养正常人毛乳头细胞的寿命且可保持其正常的生物学特性。

Objective To investigate the effect of human telomerase reverse transcriptase （hTERT） gene on normal human dermal papilla cells. Methods The hTERT gene was introduced into cultured normal human dermal papilla cells by Liposome-mediated transfection, then the positive cells were selected with G418. After the positive cells were passaged on, the integration and expression of hTERT gene were detected and the biological characteristics of the transfected cells were analyzed. Results A positive cell clone was selected, and the 35th passage of transfected dermal papilla cells was obtained so far. Expression of hTERT gene was detected on DNA, mRNA and protein levels. The transfected cells possess the biological characteristics of normal human dermal papilla cells. Conclusion The lifespan of normal human dermal papilla cells can be extended by hTERT gene transfection, and the transfected cells keep the normal biological characteristics.