论文部分内容阅读
目的 :建立永生化人牙髓干细胞系(human dental pulp stem cell,h DPSC),供口腔医学临床和基础研究使用。方法:原代培养人牙髓细胞并筛选牙髓干细胞,以慢病毒为载体,导入端粒酶反转录酶基因(h TERT)全长c DNA,经筛选得到阳性克隆。体外连续传代,应用反转录PCR(reverse transcription PCR,RT-PCR)和蛋白质印迹技术(Western blot)检测h TERT的整合和表达情况。结果 :成功地将h TERT基因转入人牙髓干细胞,得到的转化细胞端粒酶表达阳性,增殖旺盛,至今已传至第35代。结论:转导外源性h TERT基因可以导致人牙髓干细胞永生化。
Objective: To establish an immortalized human dental pulp stem cell (h DPSC) for clinical and basic research in oral medicine. Methods: Human dental pulp cells were cultured in primary culture and dental pulp stem cells were screened. The lentiviral vector was used to introduce the full length c DNA of hTERT gene and the positive clones were screened out. The cells were passaged continuously in vitro. The integration and expression of h TERT were detected by reverse transcription PCR (RT-PCR) and Western blot. Results: The hTERT gene was successfully transfected into human dental pulp stem cells and the expression of telomerase was positive. CONCLUSION: Transduction of exogenous h TERT gene can lead to immortalization of human dental pulp stem cells.