Derivation of Mesenchymal Stem Cells from Human Embryonic Stem Cells

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⁻ .