Derivation of Induced Pluripotent Stem Cells by Retroviral Gene Transduction in Mammalian Species

  • Masanori Imamura
  • Hironobu Okuno
  • Ikuo Tomioka
  • Yoshimi Kawamura
  • Zachary Yu-Ching Lin
  • Ryusuke Nakajima
  • Wado Akamatsu
  • Hirotaka James Okano
  • Yumi Matsuzaki
  • Erika Sasaki
  • Hideyuki Okano
Part of the Methods in Molecular Biology book series (MIMB, volume 925)

Abstract

Pluripotent stem cells can provide us with an enormous cell source for in vitro model systems for development. In 2006, new methodology was designed to generate pluripotent stem cells directly from somatic cells, and these cells were named induced pluripotent stem cells (iPSCs). This method consists of technically simple procedures: donor cell preparation, gene transduction, and isolation of embryonic stem cell-like colonies. The iPSC technology enables cell biologists not only to obtain pluripotent stem cells easily but also to study the reprogramming events themselves. Here, we describe the protocols to generate iPSCs from somatic origins by using conventional viral vectors. Specifically, we state the usage of three mammalian species: mouse, common marmoset, and human. As mouse iPSC donors, fibroblasts are easily prepared, while mesenchymal stem cells are expected to give rise to highly reprogrammed iPSCs efficiently. Common marmoset (Callithrix jacchus), a nonhuman primate, represents an alternative model to the usual laboratory animals. Finally, patient-specific human iPSCs give us an opportunity to examine the pathology and mechanisms of dysregulated genomic imprinting. The iPSC technology will serve as a valuable method for studying genomic imprinting, and conversely, the insights from these studies will offer valuable criteria to assess the potential of iPSCs.

Key words

Genomic imprinting Induced pluripotent stem cells Embryonic stem cells Repro-gramming Pluripotency Epigenetics Germ cells Cell culture Common marmoset Disease model 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Masanori Imamura
    • 1
  • Hironobu Okuno
    • 1
  • Ikuo Tomioka
    • 1
    • 2
  • Yoshimi Kawamura
    • 1
  • Zachary Yu-Ching Lin
    • 1
  • Ryusuke Nakajima
    • 1
  • Wado Akamatsu
    • 1
  • Hirotaka James Okano
    • 1
  • Yumi Matsuzaki
    • 1
  • Erika Sasaki
    • 1
    • 2
    • 3
  • Hideyuki Okano
    • 1
  1. 1.Department of Physiology, School of MedicineKeio UniversityTokyoJapan
  2. 2.Laboratory of Applied Developmental Biology, Marmoset Research DepartmentCentral Institute for Experimental AnimalsKawasakiJapan
  3. 3.PRESTO Japan Science and Technology AgencyTokyoJapan

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