2. Academic Validation
  3. A stress-reduced passaging technique improves the viability of human pluripotent cells

A stress-reduced passaging technique improves the viability of human pluripotent cells

  • Cell Rep Methods. 2022 Jan 24;2(2):100155. doi: 10.1016/j.crmeth.2021.100155.
Kazutoshi Takahashi 1,  Chikako Okubo 1,  Michiko Nakamura 1,  Mio Iwasaki 1,  Yuka Kawahara 1,  Tsuyoshi Tabata 1,  Yousuke Miyamoto 1,  Knut Woltjen 1,  Shinya Yamanaka 1 2 3
Affiliations

Affiliations

  • 1 Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto 606-8507, Japan.
  • 2 Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158, USA.
  • 3 Department of Anatomy, University of California, San Francisco, San Francisco, CA 94143, USA.
PMID: 35474962 DOI: 10.1016/j.crmeth.2021.100155
Abstract

Xeno-free culture systems have expanded the clinical and industrial application of human pluripotent stem cells (PSCs). However, reproducibility issues, often arising from variability during passaging steps, remain. Here, we describe an improved method for the subculture of human PSCs. The revised method significantly enhances the viability of human PSCs by lowering DNA damage and Apoptosis, resulting in more efficient and reproducible downstream applications such as gene editing and directed differentiation. Furthermore, the method does not alter PSC characteristics after long-term culture and attenuates the growth advantage of abnormal subpopulations. This robust passaging method minimizes experimental error and reduces the rate of PSCs failing quality control of human PSC research and application.

Keywords

DNA damage; human pluripotent stem cell; passage; reproducibility; viability; xeno-free culture.

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