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Reversibly immortalized hepatic progenitor cell line containing double suicide genes
A large number of functional hepatocytes is required for bioartificial liver therapy. Simian virus 40 T-antigen (SV40T) has been previously reported to improve the immortalized proliferation of primary hepatocytes to generate a sufficient number of cells; however, these long-term immortalized hepato...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
D.A. Spandidos
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108856/ https://www.ncbi.nlm.nih.gov/pubmed/30085335 http://dx.doi.org/10.3892/ijmm.2018.3803 |
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author | Fang, Shu-Yu Hu, Chao-Qun Liu, Meng-Nan Tao, Li Wang, Yi Gong, Meng-Jia He, Yun He, Tong-Chuan Bi, Yang |
author_facet | Fang, Shu-Yu Hu, Chao-Qun Liu, Meng-Nan Tao, Li Wang, Yi Gong, Meng-Jia He, Yun He, Tong-Chuan Bi, Yang |
author_sort | Fang, Shu-Yu |
collection | PubMed |
description | A large number of functional hepatocytes is required for bioartificial liver therapy. Simian virus 40 T-antigen (SV40T) has been previously reported to improve the immortalized proliferation of primary hepatocytes to generate a sufficient number of cells; however, these long-term immortalized hepatocytes may induce further malignant transformation in vivo. In the present study, the SV40T immortalization gene and two suicide genes, herpes simplex virus thymidine kinase (HSV-tk) and cytosine deaminase (CD), were transducted into primary hepatocytes to construct a novel type of Cre/LoxP-mediated reversible immortalized hepatocyte line. Polymerase chain reaction analysis and western blotting confirmed that the SV40T, HSV-tk and CD genes were successfully inserted into hepatic progenitor cells and their expression was controlled by Cre/LoxP recombination. Total removal of SV40T could be achieved via the ganciclovir (GCV)/HSV-tk suicide system. Cells maintained their biosafety in vivo with CD gene expression and 5-fluoro-cytosine (5-FC) induced cell death. Following transplantation into the carbon tetrachloride (CCl(4)) model group, the majority of cells had survived after 14 days post-implantation and a number of the cells had transported into the liver parenchyma. When compared with the CCl(4) model group, the transplanted cells repaired the liver biochemical index and pathological structure markedly. Thus, the present study reports a novel reversible immortalized hepatocyte with double suicide genes, which exhibited the cellular phenotype and recovery function of normal liver cells. This method maximally guaranteed the biological safety of immortalized hepatocytes for in vivo application, providing a reliable, safe and ideal cell material for the artificial liver technique. |
format | Online Article Text |
id | pubmed-6108856 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | D.A. Spandidos |
record_format | MEDLINE/PubMed |
spelling | pubmed-61088562018-08-27 Reversibly immortalized hepatic progenitor cell line containing double suicide genes Fang, Shu-Yu Hu, Chao-Qun Liu, Meng-Nan Tao, Li Wang, Yi Gong, Meng-Jia He, Yun He, Tong-Chuan Bi, Yang Int J Mol Med Articles A large number of functional hepatocytes is required for bioartificial liver therapy. Simian virus 40 T-antigen (SV40T) has been previously reported to improve the immortalized proliferation of primary hepatocytes to generate a sufficient number of cells; however, these long-term immortalized hepatocytes may induce further malignant transformation in vivo. In the present study, the SV40T immortalization gene and two suicide genes, herpes simplex virus thymidine kinase (HSV-tk) and cytosine deaminase (CD), were transducted into primary hepatocytes to construct a novel type of Cre/LoxP-mediated reversible immortalized hepatocyte line. Polymerase chain reaction analysis and western blotting confirmed that the SV40T, HSV-tk and CD genes were successfully inserted into hepatic progenitor cells and their expression was controlled by Cre/LoxP recombination. Total removal of SV40T could be achieved via the ganciclovir (GCV)/HSV-tk suicide system. Cells maintained their biosafety in vivo with CD gene expression and 5-fluoro-cytosine (5-FC) induced cell death. Following transplantation into the carbon tetrachloride (CCl(4)) model group, the majority of cells had survived after 14 days post-implantation and a number of the cells had transported into the liver parenchyma. When compared with the CCl(4) model group, the transplanted cells repaired the liver biochemical index and pathological structure markedly. Thus, the present study reports a novel reversible immortalized hepatocyte with double suicide genes, which exhibited the cellular phenotype and recovery function of normal liver cells. This method maximally guaranteed the biological safety of immortalized hepatocytes for in vivo application, providing a reliable, safe and ideal cell material for the artificial liver technique. D.A. Spandidos 2018-10 2018-08-02 /pmc/articles/PMC6108856/ /pubmed/30085335 http://dx.doi.org/10.3892/ijmm.2018.3803 Text en Copyright: © Fang et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. |
spellingShingle | Articles Fang, Shu-Yu Hu, Chao-Qun Liu, Meng-Nan Tao, Li Wang, Yi Gong, Meng-Jia He, Yun He, Tong-Chuan Bi, Yang Reversibly immortalized hepatic progenitor cell line containing double suicide genes |
title | Reversibly immortalized hepatic progenitor cell line containing double suicide genes |
title_full | Reversibly immortalized hepatic progenitor cell line containing double suicide genes |
title_fullStr | Reversibly immortalized hepatic progenitor cell line containing double suicide genes |
title_full_unstemmed | Reversibly immortalized hepatic progenitor cell line containing double suicide genes |
title_short | Reversibly immortalized hepatic progenitor cell line containing double suicide genes |
title_sort | reversibly immortalized hepatic progenitor cell line containing double suicide genes |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6108856/ https://www.ncbi.nlm.nih.gov/pubmed/30085335 http://dx.doi.org/10.3892/ijmm.2018.3803 |
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