Human telomerase reverse transcriptase-mediated effects on biological characteristics of rat fetal liver stem cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies have shown that stem cels transfected with human telomerase reverse transcriptase (hTERT) gene can stably express high-level telomerase activity and strengthen cel proliferation, which lays the foundation to establish geneticaly engineered immortalized stem cel lines. OBJECTIVE:To explore the effects of hTERT transfection on proliferation and cel cycle of rat fetal liver stem cels in vitro. METHODS:Rat fetal liver stem cels culturedin vitro were transfected by recombinant adeno-associated virus carrying hTERT genes. RT-PCR and western blot assay were used to detect the expression of hTERT mRNA and protein, respectively. Cel Counting Kit-8 method and cel growth curve were used to detect cel growth and proliferation. Changes in cel cycle distribution were determined by flow cytometry. RESULTS AND CONCLUSION:Compared with the control group and empty virus group, in the hTERT infection group, hTERT expressed at gene and protein levels, the growth rate of the cels increased, and the number of cels at G0/G1 phase and S phase was decreased and increased, respectively. The results show that recombinant adeno-associated virus as a vector of hTERT gene used for gene transfection can promote the in vitro proliferation of rat fetal liver stem cels and play an optimal role in cel culture.