Culture, Adaptation, and Expansion of Pluripotent Stem Cells

Methods Mol Biol. 2017;1590:139-150. doi: 10.1007/978-1-4939-6921-0_10.

Abstract

The ability of human pluripotent stem cells (hPSCs) to proliferate indefinitely in culture while maintaining their pluripotent properties makes them a powerful tool for use in research, and provides tremendous potential for diagnostic testing, and therapeutic application. Success in these areas, however, is dependent on the ability to effectively expand them in long-term culture while preserving their distinct nature. Contained in this chapter are detailed protocols for the feeder-independent culture and expansion of hPSCs using mTeSR1 medium and Matrigel matrix, and guidelines for the successful transfer of those cells to alternative platforms. These protocols have been used widely by laboratories around the world to successfully expand hPSCs for long-term culture while maintaining their undifferentiated, pluripotent state.

Keywords: Feeder-independent culture; Passaging; Pluripotent stem cell culture; Thawing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation / physiology
  • Cell Line
  • Cell Proliferation / physiology
  • Collagen / metabolism
  • Culture Media / metabolism
  • Drug Combinations
  • Feeder Cells / cytology
  • Feeder Cells / metabolism
  • Humans
  • Laminin / metabolism
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Proteoglycans / metabolism

Substances

  • Culture Media
  • Drug Combinations
  • Laminin
  • Proteoglycans
  • matrigel
  • Collagen