Construction of immortalized human liver cells with SV40T/HSV-tk gene

Objective To provide safe and effective artificial liver human liver cells by constructing immortalized normal adult liver cells.Methods Artificial liver human liver cells were transfected and packaged to obtain retrovirus which was used to infect the normal adult liver cells HL-7702.Liver cells with the HSV-tk gene were detected by ganciclovir toxicity test.SV40T mRNA-transfected liver cells were detected by RT-PCR.SV40T gene expression in transfected liver cells was detected by Western blot with immunofluorescence staining.Results Ganciclovir toxicity test showed that the transfected liver cells survived well while the untransfected liver cells died.Immortalized SV40T mRNA was detected by RT-PCR in the transfected liver cells,while SV40T protein was detected by Western blot in the transfected liver cells with immunofluorescence staining.Conclusion SV40T and HSV-tk genes can be detected in transfected normal adult liver cells and express their associated proteins.