Reprogramming hepatocellular carcinoma cell Huh7 into pluripotent stem-like cells
10.3724/SP.J.1008.2012.00603
- Author:
Jian ZHANG
1
Author Information
1. Department of Cell Biology, School of Biology and Basic Medical Sciences, Medical College, Soochow University
- Publication Type:Journal Article
- Keywords:
Hepatocellular carcinoma;
Induced pluripotent stem cells;
Lentivirus;
Reprogramming
- From:
Academic Journal of Second Military Medical University
2012;33(6):603-607
- CountryChina
- Language:Chinese
-
Abstract:
Objective To reprogram human hepatocellular carcinoma cell Huh7 into pluripotent stem cells. Methods Four recombinant lentiviruses individually carrying Oct4, Sox2, Nanog, and Lin28 were constructed and used to co-infect Huh7 cells in vitro. Post infection, the obtained pluripotent stem-like cells were identified by Alkaline phosphatase staining, immunofluorescence assay, quantitative-PCR and immunohistochemistry. The differentiation capability of the cells was examined by teratogencity test. Results Pluripotent stem-like cell colonies (iHuh7) were observed in cultured Huh7 cells after lentivirus-based induction. These colonies were positive for alkaline phosphatase staining and immunofluorescence assay showed expression of pluripotent factors: Oct4 and TRA-1-60. Quantitative-PCR indicated that several endogenous pluripotency-associated genes and stem cell-specific microRNAs were highly expressed in these pluripotent stem-like cells. In vivo differentiation test showed that iHuh7 cells could lead to teratogencity. Conclusion Huh7 cells can be induced into pluripotent stem-like cells mediated by lentiviruses individually carrying Oct4, Sox2, Nanog, and Lin28.
