Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions

Masafumi Katayama, Takashi Hirayama, Tohru Kiyono, Manabu Onuma, Tetsuya Tani, Satoru Takeda, Katsuhiko Nishimori, Tomokazu Fukuda

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

The cellular conditions required to establish induced pluripotent stem cells (iPSCs), such as the number of reprogramming factors and/or promoter selection, differ among species. The establishment of iPSCs derived from cells of previously unstudied species therefore requires the extensive optimization of programming conditions, including promoter selection and the optimal number of reprogramming factors, through a trial-and-error approach. While the four Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc are sufficient for iPSC establishment in mice, we reported previously that six reprogramming factors were necessary for the creation of iPSCs from primary prairie vole-derived cells. Further to this study, we now show detailed data describing the optimization protocol we developed in order to obtain iPSCs from immortalized prairie vole-derived fibroblasts. Immortalized cells can be very useful tools in the optimization of cellular reprogramming conditions, as cellular senescence is known to dramatically decrease the efficiency of iPSC establishment. The immortalized prairie vole cells used in this optimization were designated K4DT cells as they contained mutant forms of CDK4, cyclin D, and telomerase reverse transcriptase (TERT). We show that iPSCs derived from these immortalized cells exhibit the transcriptional silencing of exogenous reprogramming factors while maintaining pluripotent cell morphology. There were no observed differences between the iPSCs derived from primary and immortalized prairie vole fibroblasts. Our data suggest that cells that are immortalized with mutant CDK4, cyclin D, and TERT provide a useful tool for the determination of the optimal conditions for iPSC establishment.

Original languageEnglish
Pages (from-to)311-318
Number of pages8
JournalJournal of Reproduction and Development
Volume63
Issue number3
DOIs
Publication statusPublished - 2017

Keywords

  • Cellular senescence
  • Immortalized cells
  • Induced pluripotent stem cell (iPSC)
  • Pluripotency
  • Prairie vole

ASJC Scopus subject areas

  • Animal Science and Zoology

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