Sphingosine-1-Phosphate pp 167-175

Part of the Methods in Molecular Biology book series (MIMB, volume 874)

Maintenance of Human Embryonic Stem Cells by Sphingosine-1-Phosphate and Platelet-Derived Growth Factor

  • Raymond C. B. Wong
  • Martin F. Pera
  • Alice Pébay
Protocol

Abstract

Embryonic stem cells are pluripotent and capable of indefinite self-renewal in vitro. Human embryonic stem cells (hESC) have generally been cultivated on feeder layers of primary mouse embryonic fibroblasts (MEF) in media supplemented with fetal calf serum (FCS). However, serum contains a wide variety of biologically active compounds that might adversely affect hESC growth and differentiation. Thus, cultivation of stem cells in FCS complicates experimental approaches to define the intracellular mechanisms required for hESC maintenance. This chapter describes the serum-free maintenance of hESC in culture by addition of sphingosine-1-phosphate (S1P) and platelet-derived growth factor (PDGF). This complete protocol provides a chemically defined serum-free system that is advantageous for studying signaling pathways involved in hESC pluripotency.

Key words

Human embryonic stem cells Platelet-derived growth factor Serum-free medium Sphingosine-1-phosphate 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Raymond C. B. Wong
    • 1
  • Martin F. Pera
    • 2
    • 3
  • Alice Pébay
    • 4
  1. 1.Department of Biological ChemistryUniversity of California IrvineIrvineUSA
  2. 2.Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell ResearchUniversity of Southern CaliforniaLos AngelesUSA
  3. 3.Centre for NeuroscienceThe University of MelbourneParkvilleAustralia
  4. 4.Department of Pharmacology, Centre for NeuroscienceThe University of MelbourneParkvilleAustralia

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