Low-molecular-weight inhibitors of cell differentiation enable efficient growth of mouse iPS cells under feeder-free conditions

Kenichiro Donai, Akane Inagaki, Kyoung Ha So, Kengo Kuroda, Hideko Sone, Masayuki Kobayashi, Katsuhiko Nishimori, Tomokazu Fukuda

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)


Embryonic stem cells and induced pluripotent stem (iPS) cells are usually maintained on feeder cells derived from mouse embryonic fibroblasts (MEFs). In recent years, the cell culture of iPS cells under serum- and feeder-free conditions is gaining attention in overcoming the biosafety issues for clinical applications. In this study, we report on the use of multiple small-molecular inhibitors (i.e., CHIR99021, PD0325901, and Thiazovivin) to efficiently cultivate mouse iPS cells without feeder cells in a chemically-defined and serum-free condition. In this condition, we showed that mouse iPS cells are expressing the Nanog, Oct3/4, and SSEA-1 pluripotent markers, indicating that the culture condition is optimized to maintain the pluripotent status of iPS cells. Without these small-molecular inhibitors, mouse iPS cells required the adaptation period to start the stable cell proliferation. The application of these inhibitors enabled us the shortcut culture method for the cellular adaptation. This study will be useful to efficiently establish mouse iPS cell lines without MEF-derived feeder cells.

Original languageEnglish
Pages (from-to)191-197
Number of pages7
Issue number2
Publication statusPublished - 2015 Mar 18


  • Cell culture condition
  • Feeder-free
  • Induced pluripotent stem cells
  • Low-molecular-weight compounds
  • Serum-free

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Clinical Biochemistry
  • Cell Biology


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