[Transfecing hTRT into normal human oral keratinocytes by liposome]
Hai-Wen Zhou1, Wen-Jun Yang, Ke-Ming Wang
1Department of Oral Medicine, Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China. haiwen39@hotmail.com
Purpose:
To introduce human telomerase reverse transcriptase (hTRT) into normal human oral keratinocytes(OKC) to expand their life-span.
Methods:
The pEGFP-hTRT was transferred into OKCs by lipofecAMINE2000 and transferring(TF). Expression of hTERT in transfectants was examined by fluorescence microscopy, laser confocal microscopy. Transfection efficiency was examined by flow cytometer. Exogenous hTERT gene was detected by RT-PCR. The transfeced cells were selected by G418 for stable expression.
Results:
OKCs were transferred with pEGFP-hTRT by lipofecAMINE2000 and TF. The transfection efficiency was 15.34%. Exogenous hTERT gene could be detected by RT-PCR. The transfeced cells exhibited transient expression of exogenous hTERT gene.
Conclusions:
The recombinant vector pEGFP-hTRT can be transfected into OKC by liposome. TF can improve transfection efficiency. Transient expression of exogenous hTERT is possible in oral keratinocytes.
