Derivation of mesenchymal stem cells from human embryonic stem cells

Methods Mol Biol. 2011;690:175-82. doi: 10.1007/978-1-60761-962-8_12.

Abstract

Mesenchymal stem cells (MSCs) have been isolated from many tissues including differentiating human embryonic stem cells (hESCs). Derivation of MSCs from hESCs consists of two major steps: differentiation and isolation. In our hands, differentiation of hESCs towards MSC-enriched culture can be induced by trypsinizing hESCs into single cells and plating them on gelatin-coated plates in a culture condition that enhances survival of hESC-derived MSCs and not hESCs. The trypsinized hESCs were grown with feeder support and the medium was supplemented with basic fibroblast growth factor (FGF2) and platelet-derived growth factor (PDGF)-AB. A highly enriched MSC culture could be obtained by repeated passaging by trypsinization. The enriched MSC cultures could be further purified by limiting dilution or FACS sorting for CD105(+) or CD73(+) and CD24(-).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / metabolism
  • CD24 Antigen / metabolism
  • Cell Culture Techniques / methods*
  • Cell Line
  • Cell Shape
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Endoglin
  • Freezing
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / metabolism
  • Receptors, Cell Surface / metabolism

Substances

  • Antigens, CD
  • CD24 Antigen
  • ENG protein, human
  • Endoglin
  • Receptors, Cell Surface