Construction and evaluation of the SV40-LT and hTERT overexpression MDCK cell lines

Abstract

Research on establishing continuous cell line (CCL) has gained much attention recently, especially for its promising usage as an alternative substrate for vaccine production. In order to set up a ready-for-use protocol for potentially immortalizing animal originated cells, this study used MDCK as the pilot cells to explore the method using hTERT and SV40-LT genes transfected through liposome and lentivirus. Results showed that hTERT transfection with liposome and lentivirus, and SV40-LT with lentivirus could be successfully expressed in MDCK cells. Transfection efficiency of lentivirus was higher than liposome, and SV40-LT was easier to induce cell proliferation than hTERT. No mutation occurred during the passage and the original phenotype was maintained. The study provided a reference and potential method for the subsequent immortalization of primary cells.