Differentiation of multipotent mesenchymal precursors and skeletal myoblasts from human embryonic stem cells.

This unit describes a protocol for the derivation of multipotent mesenchymal precursors from human embryonic stem cells (hESCs). hESCs cultured at low density in the presence of a chemically defined serum-free medium are induced to adopt an endomesodermal fate and later a mesenchymal phenotype. FACS sorting for the surface antigen CD73 is used to purify mesenchymal precursors able to differentiate into fat, bone, cartilage, and skeletal muscle cells. Enrichment in mesenchymal precursors with a myogenic potential is achieved via an additional FACS sorting for the embryonic skeletal muscle surface marker N-CAM.