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

Overview

Journal Exp Anim

Date 2015 Oct 27

PMID 26496927

Citations 11

Authors

Masafumi Katayama

Tohru Kiyono

Kengo Horie

Takashi Hirayama

Takahiro Eitsuka

Kengo Kuroda

Kenichiro Donai

Shizu Hidema

Katsuhiko Nishimori

Tomokazu Fukuda

Affiliations

Soon will be listed here.

Abstract

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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