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Establishment of an immortalized cell line derived from the prairie vole via lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and telomerase reverse transcriptase
The prairie vole (Microtus ochrogaster) shows social behaviors such as monogamy and parenting of infants with pair bonding. These social behaviors are specific to the prairie vole and have not been observed in other types of voles, such as mountain voles. Although the prairie vole has several unique...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Japanese Association for Laboratory Animal Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4783654/ https://www.ncbi.nlm.nih.gov/pubmed/26496927 http://dx.doi.org/10.1538/expanim.15-0061 |
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author | Katayama, Masafumi Kiyono, Tohru Horie, Kengo Hirayama, Takashi Eitsuka, Takahiro Kuroda, Kengo Donai, Kenichiro Hidema, Shizu Nishimori, Katsuhiko Fukuda, Tomokazu |
author_facet | Katayama, Masafumi Kiyono, Tohru Horie, Kengo Hirayama, Takashi Eitsuka, Takahiro Kuroda, Kengo Donai, Kenichiro Hidema, Shizu Nishimori, Katsuhiko Fukuda, Tomokazu |
author_sort | Katayama, Masafumi |
collection | PubMed |
description | The prairie vole (Microtus ochrogaster) shows social behaviors such as monogamy and parenting of infants with pair bonding. These social behaviors are specific to the prairie vole and have not been observed in other types of voles, such as mountain voles. Although the prairie vole has several unique characteristics, an in vitro cell culture system has not been established for this species. Furthermore, establishment of cultured cells derived from the prairie vole may be beneficial based on the three Rs (i.e., Replacement, Reduction, and Refinement) concept. Therefore, in this study, we attempted to establish an immortalized cell line derived from the prairie vole. Our previous research has shown that transduction with mutant forms of cyclin-dependent kinase 4 (CDK4), cyclin D, and telomerase reverse transcriptase (TERT) could efficiently immortalize cells from multiple species, including humans, cattle, pigs, and monkeys. Here, we introduced these three genes into prairie vole-derived muscle fibroblasts. The expression of mutant CDK4 and cyclin D proteins was confirmed by western blotting, and telomerase activity was detected in immortalized vole muscle-derived fibroblasts (VMF-K4DT cells or VMFs) by stretch PCR. Population doubling analysis showed that the introduction of mutant CDK4, cyclin D, and TERT extended the lifespan of VMFs. To the best of our knowledge, this is the first report describing the establishment of an immortalized cell line derived from the prairie vole through the expression of mutant CDK4, cyclin D, and human TERT. |
format | Online Article Text |
id | pubmed-4783654 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Japanese Association for Laboratory Animal Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-47836542016-03-10 Establishment of an immortalized cell line derived from the prairie vole via lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and telomerase reverse transcriptase Katayama, Masafumi Kiyono, Tohru Horie, Kengo Hirayama, Takashi Eitsuka, Takahiro Kuroda, Kengo Donai, Kenichiro Hidema, Shizu Nishimori, Katsuhiko Fukuda, Tomokazu Exp Anim Original The prairie vole (Microtus ochrogaster) shows social behaviors such as monogamy and parenting of infants with pair bonding. These social behaviors are specific to the prairie vole and have not been observed in other types of voles, such as mountain voles. Although the prairie vole has several unique characteristics, an in vitro cell culture system has not been established for this species. Furthermore, establishment of cultured cells derived from the prairie vole may be beneficial based on the three Rs (i.e., Replacement, Reduction, and Refinement) concept. Therefore, in this study, we attempted to establish an immortalized cell line derived from the prairie vole. Our previous research has shown that transduction with mutant forms of cyclin-dependent kinase 4 (CDK4), cyclin D, and telomerase reverse transcriptase (TERT) could efficiently immortalize cells from multiple species, including humans, cattle, pigs, and monkeys. Here, we introduced these three genes into prairie vole-derived muscle fibroblasts. The expression of mutant CDK4 and cyclin D proteins was confirmed by western blotting, and telomerase activity was detected in immortalized vole muscle-derived fibroblasts (VMF-K4DT cells or VMFs) by stretch PCR. Population doubling analysis showed that the introduction of mutant CDK4, cyclin D, and TERT extended the lifespan of VMFs. To the best of our knowledge, this is the first report describing the establishment of an immortalized cell line derived from the prairie vole through the expression of mutant CDK4, cyclin D, and human TERT. Japanese Association for Laboratory Animal Science 2015-10-23 2016 /pmc/articles/PMC4783654/ /pubmed/26496927 http://dx.doi.org/10.1538/expanim.15-0061 Text en ©2016 Japanese Association for Laboratory Animal Science http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. |
spellingShingle | Original Katayama, Masafumi Kiyono, Tohru Horie, Kengo Hirayama, Takashi Eitsuka, Takahiro Kuroda, Kengo Donai, Kenichiro Hidema, Shizu Nishimori, Katsuhiko Fukuda, Tomokazu Establishment of an immortalized cell line derived from the prairie vole via lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and telomerase reverse transcriptase |
title | Establishment of an immortalized cell line derived from the prairie vole via
lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and
telomerase reverse transcriptase |
title_full | Establishment of an immortalized cell line derived from the prairie vole via
lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and
telomerase reverse transcriptase |
title_fullStr | Establishment of an immortalized cell line derived from the prairie vole via
lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and
telomerase reverse transcriptase |
title_full_unstemmed | Establishment of an immortalized cell line derived from the prairie vole via
lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and
telomerase reverse transcriptase |
title_short | Establishment of an immortalized cell line derived from the prairie vole via
lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin D, and
telomerase reverse transcriptase |
title_sort | establishment of an immortalized cell line derived from the prairie vole via
lentivirus-mediated transduction of mutant cyclin-dependent kinase 4, cyclin d, and
telomerase reverse transcriptase |
topic | Original |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4783654/ https://www.ncbi.nlm.nih.gov/pubmed/26496927 http://dx.doi.org/10.1538/expanim.15-0061 |
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