Published online Sep 28, 2014. doi: 10.3748/wjg.v20.i36.13119
Revised: March 30, 2014
Accepted: June 26, 2014
Published online: September 28, 2014
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.
Core tip: It is meaningful to establish reversibly immortalized human hepatocytes which can be economically grown in tissue culture. Toward this goal, we successfully established a method for the reversible immortalization of human hepatocytes using Cre/loxP site-specific recombination, which may offer a good and safe source of hepatocytes for the bioartificial liver (BAL) system in the near future. If a sufficient number of human hepatocytes can be used, these extracorporeal devices will serve as successful “bridge-to-transplant”therapies. With the progress made in bioreactor development, the next-generation BAL system could reach the level of artificial kidney and save more patients.