TY - JOUR
T1 - 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
AU - Katayama, Masafumi
AU - Hirayama, Takashi
AU - Kiyono, Tohru
AU - Onuma, Manabu
AU - Tani, Tetsuya
AU - Takeda, Satoru
AU - Nishimori, Katsuhiko
AU - Fukuda, Tomokazu
N1 - Funding Information:
We thank Dr Shizu Hidema, Dr Kengo Kuroda, Mr Kenichiro Donai, and Mr Kengo Horie for helpful discussions. We thank Dr Larry J Young (Yerkes National Primate Research Center, Dept. of Psychiatry, Emory University) for providing the prairie voles used in these experiments. We thank Gustavo Mostoslavsky (Bos-ton University School of Medicine) for providing the STEMCCA-loxP lentiviral vector.
Publisher Copyright:
© 2017 by the Society for Reproduction and Development.
PY - 2017
Y1 - 2017
N2 - 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.
AB - 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.
KW - Cellular senescence
KW - Immortalized cells
KW - Induced pluripotent stem cell (iPSC)
KW - Pluripotency
KW - Prairie vole
UR - http://www.scopus.com/inward/record.url?scp=85021228813&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85021228813&partnerID=8YFLogxK
U2 - 10.1262/jrd.2016-164
DO - 10.1262/jrd.2016-164
M3 - Article
C2 - 28331164
AN - SCOPUS:85021228813
VL - 63
SP - 311
EP - 318
JO - Journal of Reproduction and Development
JF - Journal of Reproduction and Development
SN - 0916-8818
IS - 3
ER -