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Updated: Apr 23, 2026
Lentiviral Vector-mediated Gene Therapy of Hepatocytes Ex Vivo for Autologous Transplantation in Swine
Published on: November 4, 2018
Reversible immortalization of human hepatocytes mediated by retroviral transfer and site-specific recombination
Fan-Ying Meng1, Li Liu1, Feng-Hui Yang1
1Fan-Ying Meng, Feng-Hui Yang, Chun-You Li, Jun Liu, Division of Hepato-Biliary-Pancreatic Surgery, Institute of Organ Transplantation, Shandong Provincial Hospital Affiliated to Shandong University, Jinan 250021, Shandong Province, China.
Aim:
To establish a method for the reversible immortalization of human hepatocytes, which may offer a good and safe source of hepatocytes for practical applications.
Methods:
We successfully isolated primary human hepatocytes from surgically resected liver tissue taken from a patient with liver hemangiomas. The freshly isolated cells were then immortalized with retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40T) and hygromycin-resistance genes flanked by paired loxP recombination targets.
Results:
The freshly isolated hepatocytes with high viability (85%) were successfully immortalized using retroviral gene transfer of SV40T. SV40T in the immortalized cells was then excised by Cre/loxP site-specific recombination. This cell population exhibited the characteristics of differentiated hepatocytes.
Conclusion:
We successfully established reversibly immortalized human hepatocytes, which will provide an unlimited supply of cells for practical applications.
