Establishment of a hepatic stem cell line stably expressing human interleukin-2 / 中国免疫学杂志

ABSTRACT

Objective:Establishing the hepatic stem cell(WB-F344)line that expresses human interleukin-2 stably and effectively.Methods: We constructed the retroviral vector Plpcx-il-2 containing human interleukin-2 gene, and transfected it into PT67 packaging cells.The infectious viruses were used to infect the hepatic stem cell WB-F344,then the puromycin resistant clones were acquired after puromycin selection.The expression of IL-2 was detected using RT-PCR,immunocytochemistry, Western blot and indirect immunofluorescence, respectively.Results: The retroviral vector containing human IL-2 gene was constructed.The titer of virus in the supernatant reached 10~5 CFU/ml after transfecting PT67 packeting cells.The puromycin resistant clones were acquired after infection of WB-F344 and the cell line WB-F344/Plpcx-IL-2 expressing human interleukin-2 stably was established.Conclusion: By retroviral-mediated transfecting human interleukin-2 gene, the hepatic stem cell stably and effectively expressing interleukin-2 are obtained.This provides a good basis for further research on immuno-gene therapy of liver cancer.