Step-by-step protocols for non-viral derivation of transgene-free induced pluripotent stem cells from somatic fibroblasts of multiple mammalian species

Sho Yoshimatsu, Atsushi Yamazaki, Kazuya Edamura, Yuko Koushige, Hisashi Shibuya, Emi Qian, Tsukika Sato, Junko Okahara, Noriyuki Kishi, Toshiaki Noce, Yoshifumi Yamaguchi, Hideyuki Okano

Research output: Contribution to journalArticlepeer-review

Abstract

Potentials of immortal proliferation and unlimited differentiation into all the three germ layers and germ cells in induced pluripotent stem cells (iPSCs) render them important bioresources for in vitro reconstitution and modeling of intravital tissues and organs in various animal models, thus contributing to the elucidation of pathomechanisms, drug discovery and stem cell-based regenerative medicine. We previously reported promising approaches for deriving transgene-free iPSCs from somatic fibroblasts of multiple mammalian species by episomal vector or RNA transfection, although the respective step-by-step protocols and the combinatorial usage of these methods, which achieved high induction efficiency, have not been described in the literature so far. Here, we provide a detailed step-by-step description of these methods with critical tips and slight modifications (improvements) to previously reported methods. We also report a novel method for the establishment of iPSCs from the Syrian hamster (also known as golden hamster; Mesocricetus auratus), a unique animal model of hibernation. We anticipate this methodology will contribute to stem cell biology and regenerative medicine research.

Original languageEnglish
Pages (from-to)325-341
Number of pages17
JournalDevelopment Growth and Differentiation
Volume64
Issue number6
DOIs
Publication statusPublished - 2022 Aug

Keywords

  • induced pluripotent stem cell
  • mammalian
  • reprogramming

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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