Long-Term In Vitro Expansion of Epithelial Stem Cells Enabled by Pharmacological Inhibition of PAK1-ROCK-Myosin II and TGF-β Signaling

Chengkang Zhang, Hyung Joo Lee, Anura Shrivastava, Ruipeng Wang, Travis J. McQuiston, Sharon S. Challberg, Brian A. Pollok, Ting Wang

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

Despite substantial self-renewal capability in vivo, epithelial stem and progenitor cells located in various tissues expand for a few passages in vitro in feeder-free condition before they succumb to growth arrest. Here, we describe the EpiX method, which utilizes small molecules that inhibit PAK1-ROCK-Myosin II and TGF-β signaling to achieve over one trillion-fold expansion of human epithelial stem and progenitor cells from skin, airway, mammary, and prostate glands in the absence of feeder cells. Transcriptomic and epigenomic studies show that this condition helps epithelial cells to overcome stresses for continuous proliferation. EpiX-expanded basal epithelial cells differentiate into mature epithelial cells consistent with their tissue origins. Whole-genome sequencing reveals that the cells retain remarkable genome integrity after extensive in vitro expansion without acquiring tumorigenicity. EpiX technology provides a solution to exploit the potential of tissue-resident epithelial stem and progenitor cells for regenerative medicine. Zhang et al. screen a small-molecule collection and find that pharmacologic inhibition of TGF-β and PAK1-ROCK-Myosin II, in low calcium conditions, supports extended expansion of epithelial stem cells in 2D format. This approach enhances the potential of tissue-resident epithelial stem cells for cell therapy.

Original languageEnglish
Pages (from-to)598-610.e5
JournalCell Reports
Volume25
Issue number3
DOIs
StatePublished - Oct 16 2018

Keywords

  • PAK1/ROCK/Myosin II
  • TGF-β
  • cell culture method
  • cell therapy
  • epithelial stem and progenitor cells
  • feeder-free
  • regenerative medicine

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