关键词: 间充质干细胞, 诱导分化, 脂细胞, 成骨细胞

Abstract: Abstract:Objective To establish the method for isolation and cultivation of mesenchymal stem cells(MSCs) from human embryo in vitro,and to identify their biological characteristics. Methods Cells were separated from human embryo 5-6 weeks after digestion with 0.25% trypsin for 7-10 min,and the anchoring cells were expanded in MSCs media.Flow cytometry(FCM) was adopted to determine the immunophenotypes of MSCs,and the proliferation ability and cell cycle were etected by MTT and propidium iodide(PI) staining. Different induction conditions were used to identify the multipotent differentiation.Results MSCs were obtained from human embryo by tissue digestion. FCM result showed the immunophenotype of MSCs was positive for CD29,CD44,CD73,CD105 and CD166,and negative for CD34,CD45 and HLA-DR.The doubling time of MSCs was 23 h.Lipid droplets were found in lipo-induced MSCs，and under ossa-induced condition,the histological analysis of the cells revealed stronger expression of alkatnephosphatase and the mineralization of calcium salts was confirmed by alizarin red . Conclusion The phenotype of MSCs derived from human early embryo is coincident with adult MSCs,and these cells have the potential to differentiate into adipocytes and osteoblasts.

Key words: mesenchymal stem cells, induction and differentiation, adipocytes, osteoblasts