Abstract: Objective To establish a differentiation inducing method of mesenchymal stem cells (MSCs) from human embryonic stem cells (hESCs) using low-serum containing medium. Method The induction was initiated by culturing human embryonic stem cells in low-serum containing medium, the medium was half-changed every 3 days. 7～10 days later, cells were confluent and dissociated into single cells, then culture medium was changed to proliferation medium, cells were subcultured every 3 days. Results hESCs were cultured in feeder free condition and grew as clones with serrated edges, all clones expressed pluripotency markes Oct4, Sox2 and Nanog. After 3 days’induction, fibroblast-like cells with small volume appeared between hESCs clones. These cells grew as clusters and possessed high proliferation potential. Cells with uniform spindly morphology were obtained after 2 passage. All cells expressed mesenchymal marker-vimentin. Flowcytometry analysis showed that these cells expressed CD105, CD90, CD73, CD44 and HLA-ABC, no expression of CD34, CD45 and HLA-DR was detected. Under certain condition, these cells could differentiate into osteoblasts and fat cells. Conclusion Human embryonic stem cell derived mesenchymal stem cells (hESC-MSCs) with high purity can be obtained in a short time by using low-serum containing induction medium.

Key words: mesenchymal stem cells, embryonic stem cells, low-serum induction