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.
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.
Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan.|National Institute for Environmental Studies, Japan, Center for Environmental Biology and Ecosystem Studies, Ibaraki 305-8506, Japan.|Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Ibaraki 305-8506, Japan.
Masafumi
M
Katayama
Department of Obstetrics and Gynecology, Juntendo University, Tokyo 113-8421, Japan.
Takashi
T
Hirayama
Division of Carcinogenesis and Cancer Prevention, National Cancer Center Research Institute, Tokyo 104-0045, Japan.
Tohru
T
Kiyono
National Institute for Environmental Studies, Japan, Center for Environmental Biology and Ecosystem Studies, Ibaraki 305-8506, Japan.|Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Ibaraki 305-8506, Japan.
Manabu
M
Onuma
Laboratory of Animal Reproduction, Department of Agriculture, Kindai University, Nara 3327-204, Japan.
Tetsuya
T
Tani
Department of Obstetrics and Gynecology, Juntendo University, Tokyo 113-8421, Japan.
Satoru
S
Takeda
Graduate School of Agricultural Science, Tohoku University, Sendai 981-8555, Japan.
Katsuhiko
K
Nishimori
Wildlife Genome Collaborative Research Group, National Institute for Environmental Studies, Ibaraki 305-8506, Japan.|United Graduate School of Agricultural Sciences, Iwate University, Morioka 020-8551, Japan.
Tomokazu
T
Fukuda
D000818:Animals; D003411:Arvicolinae; D065150:Cellular Reprogramming; D005347:Fibroblasts; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells; D000090062:Kruppel-Like Factor 4; D016086:Lentivirus; D061251:Primary Cell Culture
Cellular senescence; Immortalized cells; Induced pluripotent stem cell (iPSC); Pluripotency; Prairie vole