Cre/loxP-Based Reversible Immortalization of Human Hepatocytes
An ideal alternative to the primary human hepatocytes for hepatocyte transplantation would be to use a clonal cell line that grows economically in culture and exhibits the characteristics of differentiated, nontransformed hepatocytes following transplantation. The purpose of the present studies was...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2001-05-01
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| Series: | Cell Transplantation |
| Online Access: | https://doi.org/10.3727/000000001783986558 |
| _version_ | 1829139729905352704 |
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| author | Naoya Kobayashi M.D., Ph.D. Hirofumi Noguchi Karen A. Westerman Takamasa Watanabe Toshihisa Matsumura Toshinori Totsugawa Toshiyoshi Fujiwara Philippe Leboulch Noriaki Tanaka |
| author_facet | Naoya Kobayashi M.D., Ph.D. Hirofumi Noguchi Karen A. Westerman Takamasa Watanabe Toshihisa Matsumura Toshinori Totsugawa Toshiyoshi Fujiwara Philippe Leboulch Noriaki Tanaka |
| author_sort | Naoya Kobayashi M.D., Ph.D. |
| collection | DOAJ |
| description | An ideal alternative to the primary human hepatocytes for hepatocyte transplantation would be to use a clonal cell line that grows economically in culture and exhibits the characteristics of differentiated, nontransformed hepatocytes following transplantation. The purpose of the present studies was to establish a reversibly immortalized human hepatocyte cell line. Human hepatocytes were immortalized with a retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40Tag) gene flanked by a pair of loxP recombination targets. One of the resulting clones, NKNT-3, showed morphological characteristics of liver parenchymal cells and expressed the genes of differentiated liver functions. NKNT-3 cells offered unlimited availability. After an adenoviral delivery of Cre recombinase and subsequent differential selection, efficient removal of SV40Tag from NKNT-3 cells was performed. Here we represent that elimination of the retrovirally transferred SV40Tag gene can be excised by adenovirus-mediated site-specific recombination. |
| first_indexed | 2024-12-14T19:39:24Z |
| format | Article |
| id | doaj.art-600036b49e2d4b8d9528c2218746c5d6 |
| institution | Directory Open Access Journal |
| issn | 0963-6897 1555-3892 |
| language | English |
| last_indexed | 2024-12-14T19:39:24Z |
| publishDate | 2001-05-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Cell Transplantation |
| spelling | doaj.art-600036b49e2d4b8d9528c2218746c5d62022-12-21T22:49:44ZengSAGE PublishingCell Transplantation0963-68971555-38922001-05-011010.3727/000000001783986558Cre/loxP-Based Reversible Immortalization of Human HepatocytesNaoya Kobayashi M.D., Ph.D.0Hirofumi Noguchi1Karen A. Westerman2Takamasa Watanabe3Toshihisa Matsumura4Toshinori Totsugawa5Toshiyoshi Fujiwara6Philippe Leboulch7Noriaki Tanaka8First Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanFirst Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanHarvard-Massachusetts Institute of Technology, Division of Health Sciences and Technology, Cambridge, MA 02139First Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanFirst Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanFirst Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanFirst Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanHarvard-Massachusetts Institute of Technology, Division of Health Sciences and Technology, Cambridge, MA 02139First Department of Surgery, Okayama University Medical School, 2-5-1 Shikata-cho, Okayama 700-8558, JapanAn ideal alternative to the primary human hepatocytes for hepatocyte transplantation would be to use a clonal cell line that grows economically in culture and exhibits the characteristics of differentiated, nontransformed hepatocytes following transplantation. The purpose of the present studies was to establish a reversibly immortalized human hepatocyte cell line. Human hepatocytes were immortalized with a retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40Tag) gene flanked by a pair of loxP recombination targets. One of the resulting clones, NKNT-3, showed morphological characteristics of liver parenchymal cells and expressed the genes of differentiated liver functions. NKNT-3 cells offered unlimited availability. After an adenoviral delivery of Cre recombinase and subsequent differential selection, efficient removal of SV40Tag from NKNT-3 cells was performed. Here we represent that elimination of the retrovirally transferred SV40Tag gene can be excised by adenovirus-mediated site-specific recombination.https://doi.org/10.3727/000000001783986558 |
| spellingShingle | Naoya Kobayashi M.D., Ph.D. Hirofumi Noguchi Karen A. Westerman Takamasa Watanabe Toshihisa Matsumura Toshinori Totsugawa Toshiyoshi Fujiwara Philippe Leboulch Noriaki Tanaka Cre/loxP-Based Reversible Immortalization of Human Hepatocytes Cell Transplantation |
| title | Cre/loxP-Based Reversible Immortalization of Human Hepatocytes |
| title_full | Cre/loxP-Based Reversible Immortalization of Human Hepatocytes |
| title_fullStr | Cre/loxP-Based Reversible Immortalization of Human Hepatocytes |
| title_full_unstemmed | Cre/loxP-Based Reversible Immortalization of Human Hepatocytes |
| title_short | Cre/loxP-Based Reversible Immortalization of Human Hepatocytes |
| title_sort | cre loxp based reversible immortalization of human hepatocytes |
| url | https://doi.org/10.3727/000000001783986558 |
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