RRC ID 49216
著者 Katayama M, Hirayama T, Kiyono T, Onuma M, Tani T, Takeda S, Nishimori K, Fukuda T.
タイトル 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.
ジャーナル J Reprod Dev
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.
巻・号 63(3)
ページ 311-318
公開日 2017-6-21
DOI 10.1262/jrd.2016-164
PMID 28331164
PMC PMC5481634
MeSH Animals Arvicolinae Cellular Reprogramming* Fibroblasts Genetic Vectors HEK293 Cells Humans Induced Pluripotent Stem Cells* Kruppel-Like Factor 4 Lentivirus Primary Cell Culture
IF 1.735
引用数 6
WOS 分野 AGRICULTURE, DAIRY & ANIMAL SCIENCE REPRODUCTIVE BIOLOGY
リソース情報
遺伝子材料 pCAG-HIVgp (RDB04394) pCMV-VSV-G-RSV-Rev (RDB04393)