Human telomerase reverse transcriptase gene-transfected effects on biological characteristics of Schwann cells / 中国组织工程研究

ABSTRACT

BACKGROUND:Studies have shown that genetical y modified Schwann cel s can survive for a longer time in vivo, and promote nerve regeneration and functional recovery. OBJECTIVE:To transfect human telomerase reverse transcriptase (hTERT) gene into rat Schwann cel s cultured in vitro via PLXSN vector, and to detect the telomerase activity and biological characteristics of Schwann cel s. METHODS:Schwann cel s from Wistar rats were cultured in vitro and transfected by PLXSN vector with (hTERT group) or without hTERT (empty vector group). Normal Schwann cel s were selected as control group. RT-PCR and western blot methods were used to detect the hTERT protein and mRNA levels in Schwann cel s, and flow cytometry was used to measure the cel cycle distribution. Cel growth was observed by cel growth curve and MTT colorimetric method. RESULTS AND CONCLUSION:At 48 hours after transfection, the mRNA and protein expressions of hTERT were remarkably seen in Schwann cel s. Compared with the control and empty vector groups, the cel s grew faster, the number of cel s at G 0/G 1 Schwann cel s cultured in vitro. phase was reduced, but the number of S phase cel s was increased in the hTERT group (P<0.05). These findings indicate that PLXSN vector-mediated hTERT transfection of Schwann cel s can significantly improve the activity of telomerase in Schwann cel s as wel as promote the proliferation of Schwann cells cultured in vitro.