Dedifferentiation of conditionally immortalized hepatocytes with long-term in vitro passage

Dedifferentiation of conditionally immortalized hepatocytes with long-term in vitro passage

Play all audios:

Loading...

ABSTRACT The rat hepatocytes were immortalized using a temperature-sensitive mutant of SV40 large T antigen (tsT) to develop as a possible substitute for primary hepatocytes. Four rat hepatocyte lines that have been developed and maintained more than passage 50, were characterized for their cellular morphology, T antigen and p53 expression, chromosomes, liver-specific differentiation, telomerase activity and anchorage independent growth. All of four cell lines showed a typical epithelial cell morphology, but the population-doubling time became short with passage: 18 to 60%. T antigen expression was increased with passage about 3 to 65 times at permissive temperature but decreased significantly at non-permissive temperature. The expression level of p53 unchanged during passages was also decreased at non-permissive temperature. The distribution of chromosome number changed somewhat with passage. The production levels of albumin and urea in four cell lines were 2.4 to 13.0% and 7.5 to 19.9% of those produced in primary hepatocytes, respectively and were decreased to an undetectable level with passage. Telomerase activity was increased 10 fold following immortalization of cells, but anchorage independent growth of cells did not develop. These results indicate that conditionally immortalized hepatocytes become dedifferentiated with in vitro passage, which may be caused by marked chromosomal damages that occur with compulsive and continuous replications by the increment of T antigen content with passage and its sequential inhibition of p53 function. SIMILAR CONTENT BEING VIEWED BY OTHERS HUMAN CELL TRANSFORMATION BY COMBINED LINEAGE CONVERSION AND ONCOGENE EXPRESSION Article Open access 23 July 2021 IL6 SUPPORTS LONG-TERM EXPANSION OF HEPATOCYTES IN VITRO Article Open access 29 November 2022 IMMORTALIZATION OF HUMAN HEPATOCYTES FROM BILIARY ATRESIA WITH CDK4R24C, CYCLIN D1, AND TERT FOR CYTOCHROME P450 INDUCTION TESTING Article Open access 15 October 2020 ARTICLE PDF AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Internal Medicine, Kyung Hee University College of Medicine, Seoul, Korea Byung-Ho Kim Authors * Byung-Ho Kim View author publications You can also search for this author inPubMed Google Scholar * Se-Ra Sung View author publications You can also search for this author inPubMed Google Scholar * Eun-Hee Choi View author publications You can also search for this author inPubMed Google Scholar * Young-Il Kim View author publications You can also search for this author inPubMed Google Scholar * Kyeong-Jin Kim View author publications You can also search for this author inPubMed Google Scholar * Seok-Ho Dong View author publications You can also search for this author inPubMed Google Scholar * Hyo-Jong Kim View author publications You can also search for this author inPubMed Google Scholar * Young-Woon Chang View author publications You can also search for this author inPubMed Google Scholar * Joung-Il Lee View author publications You can also search for this author inPubMed Google Scholar * Rin Chang View author publications You can also search for this author inPubMed Google Scholar RIGHTS AND PERMISSIONS This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Kim, BH., Sung, SR., Choi, EH. _et al._ Dedifferentiation of conditionally immortalized hepatocytes with long-term in vitro passage. _Exp Mol Med_ 32, 29–37 (2000). https://doi.org/10.1038/emm.2000.6 Download citation * Published: 01 March 2000 * Issue Date: 01 March 2000 * DOI: https://doi.org/10.1038/emm.2000.6 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * chromosome * conditional immortalization * liverspecific differentiation * Rat hepatocytes * temperature-sensitive SV40 large T antigen * telomerase

ABSTRACT The rat hepatocytes were immortalized using a temperature-sensitive mutant of SV40 large T antigen (tsT) to develop as a possible substitute for primary hepatocytes. Four rat


hepatocyte lines that have been developed and maintained more than passage 50, were characterized for their cellular morphology, T antigen and p53 expression, chromosomes, liver-specific


differentiation, telomerase activity and anchorage independent growth. All of four cell lines showed a typical epithelial cell morphology, but the population-doubling time became short with


passage: 18 to 60%. T antigen expression was increased with passage about 3 to 65 times at permissive temperature but decreased significantly at non-permissive temperature. The expression


level of p53 unchanged during passages was also decreased at non-permissive temperature. The distribution of chromosome number changed somewhat with passage. The production levels of albumin


and urea in four cell lines were 2.4 to 13.0% and 7.5 to 19.9% of those produced in primary hepatocytes, respectively and were decreased to an undetectable level with passage. Telomerase


activity was increased 10 fold following immortalization of cells, but anchorage independent growth of cells did not develop. These results indicate that conditionally immortalized


hepatocytes become dedifferentiated with in vitro passage, which may be caused by marked chromosomal damages that occur with compulsive and continuous replications by the increment of T


antigen content with passage and its sequential inhibition of p53 function. SIMILAR CONTENT BEING VIEWED BY OTHERS HUMAN CELL TRANSFORMATION BY COMBINED LINEAGE CONVERSION AND ONCOGENE


EXPRESSION Article Open access 23 July 2021 IL6 SUPPORTS LONG-TERM EXPANSION OF HEPATOCYTES IN VITRO Article Open access 29 November 2022 IMMORTALIZATION OF HUMAN HEPATOCYTES FROM BILIARY


ATRESIA WITH CDK4R24C, CYCLIN D1, AND TERT FOR CYTOCHROME P450 INDUCTION TESTING Article Open access 15 October 2020 ARTICLE PDF AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of


Internal Medicine, Kyung Hee University College of Medicine, Seoul, Korea Byung-Ho Kim Authors * Byung-Ho Kim View author publications You can also search for this author inPubMed Google


Scholar * Se-Ra Sung View author publications You can also search for this author inPubMed Google Scholar * Eun-Hee Choi View author publications You can also search for this author inPubMed


 Google Scholar * Young-Il Kim View author publications You can also search for this author inPubMed Google Scholar * Kyeong-Jin Kim View author publications You can also search for this


author inPubMed Google Scholar * Seok-Ho Dong View author publications You can also search for this author inPubMed Google Scholar * Hyo-Jong Kim View author publications You can also search


for this author inPubMed Google Scholar * Young-Woon Chang View author publications You can also search for this author inPubMed Google Scholar * Joung-Il Lee View author publications You


can also search for this author inPubMed Google Scholar * Rin Chang View author publications You can also search for this author inPubMed Google Scholar RIGHTS AND PERMISSIONS This is an


Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted


non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Kim, BH., Sung,


SR., Choi, EH. _et al._ Dedifferentiation of conditionally immortalized hepatocytes with long-term in vitro passage. _Exp Mol Med_ 32, 29–37 (2000). https://doi.org/10.1038/emm.2000.6


Download citation * Published: 01 March 2000 * Issue Date: 01 March 2000 * DOI: https://doi.org/10.1038/emm.2000.6 SHARE THIS ARTICLE Anyone you share the following link with will be able to


read this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing


initiative KEYWORDS * chromosome * conditional immortalization * liverspecific differentiation * Rat hepatocytes * temperature-sensitive SV40 large T antigen * telomerase