Live Cell Monitoring and Enrichment of Stem Cell-Derived β Cells Using Intracellular Zinc Content as a Population Marker

Curr Protoc Stem Cell Biol. 2019 Dec;51(1):e99. doi: 10.1002/cpsc.99.

Abstract

Our laboratory and others have developed protocols to generate glucose-responsive stem cell-derived β cells in vitro. The cells resulting from these protocols could supplement or replace the use of human cadaveric islets for cell-based therapy for diabetes. The combination of an unlimited supply of pluripotent stem cell-derived β cells and gene-editing approaches will facilitate numerous in vitro studies not possible with cadaveric islets. Here, we describe a protocol for fluorescent labeling and isolation of stem cell-derived β cells. This purification of SC-β cells is based on intracellular zinc content and is a simple method to complement other approaches for generating and assaying these cells. © 2019 The Authors. Basic Protocol: Fluorescent labeling and isolation of stem cell-derived β cells.

Keywords: SC-β cell enrichment; intracellular zinc content; stem cell-derived β (SC-β) cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods
  • Humans
  • Insulin-Secreting Cells / cytology*
  • Staining and Labeling / methods*
  • Zinc / metabolism*

Substances

  • Zinc