Induction Of Pluripotent Stem Cells From Hepatocellular Carcinoma Cell Lines

Induced pluripotent stem cells (iPS cells) are derived from somatic cells that regainpluripotency by the nuclear programming with exogenous factors. iPS cells haveimmense potential applications in establishing disease models and understandingdisease mechanisms, cell therapies, etc. avoiding both immunologic and ethical barriersto their use. iPS–based disease models have made great achievements. However, it isnecessary to overcome challenges such as low reprogramming efficiency and risk due totumorigenicity.Here，we reprogrammed the human hepatocellular carcinoma cell Huh7intopluripotent stem cells. Huh7is one of the immortalized human hepatocellular carcinomacell that cultured in vitro. We used four recombinant lentivirus individually carryingOct4, Sox2, Nanog, and Lin28were constructed and used to co-infect Huh7cells invitro. Post infection, the obtained pluripotent stem-like cells were identified by Alkalinephosphatase staining, immunofluorescence assay, quantitative-PCR andimmunohistochemistry. After lentivirus-based induction, pluripotent stem-like cellcolonies could be observed in cultured Huh7cells. These colonies showed positive inalkaline phosphatase staining and were expressing the pluripotent factors Oct4andTRA-1-60.The results of the quantitative-PCR indicated that several endogenouspluripotency-associated genes and stem cell-specific microRNAs were highlyexpressing in these pluripotent stem-like cells. So,we concluded that Huh7cells couldbe induced into pluripotent stem-like cells,which are mediated by lentivirus individuallycarrying Oct4, Sox2, Nanog, and Lin28.