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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...

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Autores principales: Katayama, Masafumi, Kiyono, Tohru, Horie, Kengo, Hirayama, Takashi, Eitsuka, Takahiro, Kuroda, Kengo, Donai, Kenichiro, Hidema, Shizu, Nishimori, Katsuhiko, Fukuda, Tomokazu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Association for Laboratory Animal Science 2015
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.
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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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