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The Generation and Maintenance of Rat Induced Pluripotent Stem Cells

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Stem Cells and Tissue Repair

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1210))

Abstract

This chapter describes a newly developed method for generating and maintaining rat induced pluripotent stem cells (riPSCs). We first provide a detailed protocol for the generation of lentiviral vector carrying three reprogramming factors to produce high-quality riPSCs. This technique allows reprogramming of rat somatic cells to ground state with germ-line competence. Subsequently, we elaborate a detailed protocol for the generation of riPSCs from rat embryonic fibroblast (REF). Finally, the protocols for the optimal culture conditions of riPSCs and preparation of frozen stock are described. We also outline the advantages of generating riPSCs.

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References

  1. Takahashi K, Yamanaka S (2006) Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126(4):663–676. doi:10.1016/j.cell.2006.07.024, S0092-8674(06)00976-7 [pii]

    Article  CAS  PubMed  Google Scholar 

  2. Okita K, Ichisaka T, Yamanaka S (2007) Generation of germLine-competent induced pluripotent stem cells. Nature 448(7151):313–317. doi:10.1038/nature05934, nature05934 [pii]

    Article  CAS  PubMed  Google Scholar 

  3. Li P, Tong C, Mehrian-Shai R, Jia L, Wu N, Yan Y, Maxson RE, Schulze EN, Song H, Hsieh CL, Pera MF, Ying QL (2008) GermLine competent embryonic stem cells derived from rat blastocysts. Cell 135(7):1299–1310. doi:10.1016/j.cell.2008.12.006, S0092-8674(08)01567-5 [pii]

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Buehr M, Meek S, Blair K, Yang J, Ure J, Silva J, McLay R, Hall J, Ying QL, Smith A (2008) Capture of authentic embryonic stem cells from rat blastocysts. Cell 135(7):1287–1298. doi:10.1016/j.cell.2008.12.007, S0092-8674(08)01568-7 [pii]

    Article  CAS  PubMed  Google Scholar 

  5. Ying QL, Wray J, Nichols J, Batlle-Morera L, Doble B, Woodgett J, Cohen P, Smith A (2008) The ground state of embryonic stem cell self-renewal. Nature 453(7194):519–523. doi:10.1038/nature06968, nature06968 [pii]

    Article  CAS  PubMed  Google Scholar 

  6. Li W, Wei W, Zhu S, Zhu J, Shi Y, Lin T, Hao E, Hayek A, Deng H, Ding S (2009) Generation of rat and human induced pluripotent stem cells by combining genetic reprogramming and chemical inhibitors. Cell Stem Cell 4(1):16–19. doi:10.1016/j.stem.2008.11.014, S1934-5909(08)00616-4 [pii]

    Article  PubMed  Google Scholar 

  7. Hamanaka S, Yamaguchi T, Kobayashi T, Kato-Itoh M, Yamazaki S, Sato H, Umino A, Wakiyama Y, Arai M, Sanbo M, Hirabayashi M, Nakauchi H (2011) Generation of germLine-competent rat induced pluripotent stem cells. PLoS One 6(7):e22008. doi:10.1371/journal.pone.0022008, PONE-D-11-06622 [pii]

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. Yamaguchi T, Hamanaka S, Kamiya A, Okabe M, Kawarai M, Wakiyama Y, Umino A, Hayama T, Sato H, Lee YS, Kato-Itoh M, Masaki H, Kobayashi T, Yamazaki S, Nakauchi H (2012) Development of an all-in-one inducible lentiviral vector for gene specific analysis of reprogramming. PLoS One 7(7):e41007. doi:10.1371/journal.pone.0041007, PONE-D-12-06326 [pii]

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. Okabe M, Otsu M, Ahn DH, Kobayashi T, Morita Y, Wakiyama Y, Onodera M, Eto K, Ema H, Nakauchi H (2009) Definitive proof for direct reprogramming of hematopoietic cells to pluripotency. Blood 114(9):1764–1767. doi:10.1182/blood-2009-02-203695, blood-2009-02-203695 [pii]

    Article  CAS  PubMed  Google Scholar 

  10. Nishimura K, Sano M, Ohtaka M, Furuta B, Umemura Y, Nakajima Y, Ikehara Y, Kobayashi T, Segawa H, Takayasu S, Sato H, Motomura K, Uchida E, Kanayasu-Toyoda T, Asashima M, Nakauchi H, Yamaguchi T, Nakanishi M (2011) Development of defective and persistent Sendai virus vector: a unique gene delivery/expression system ideal for cell reprogramming. J Biol Chem 286(6):4760–4771. doi:10.1074/jbc.M110.183780, M110.183780 [pii]

    Article  CAS  PubMed Central  PubMed  Google Scholar 

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Correspondence to Tomoyuki Yamaguchi .

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Yamaguchi, T., Hamanaka, S., Nakauchi, H. (2014). The Generation and Maintenance of Rat Induced Pluripotent Stem Cells. In: Kioussi, C. (eds) Stem Cells and Tissue Repair. Methods in Molecular Biology, vol 1210. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1435-7_11

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  • DOI: https://doi.org/10.1007/978-1-4939-1435-7_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1434-0

  • Online ISBN: 978-1-4939-1435-7

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