脐带源间充质干细胞的分离培养鉴定及成骨成脂诱导分化

objective: To explore the methods for isolating , cultivation and identification mesenchymal stem cells (MSCs) from the tissue of umbilical cord , with an aim to induce osteogenic and adipogenic differentiation in vitro. Method: hUC-MSCs(human umbilical cord mesenchymal stem cell) were isolated from human umbilical cord by culture of tissue adherence. The surface of passage 3 cell surface antigen was measured by flow cytometry; The cells were induced by osteogenic and adipogenic induction medium. Results: A few cells migrated out from tissue blocks, showing a shuttle shape after 8 days of culture; The primary cells growing up to 70%~80% confluence after 14 days of culture,after subculture, they were swirling aggregation or parallel growth ; The flow cytometry analysis revealed that CD29,CD105 were highly expressed on the surface of passages 3 cells, but the expression was negative for CD34, HLA-DR . After 20 days induction, alizarin red staining were positive, it proved the formation of calcium node, the liquid vacuoles were also detected by oil red O staining after culture induction of adipogenic differentiation 25 days later. Conclusion: Through morphology, flow cytometry immunizing, the osteogenic adipogenic differentiation, it was recognized as hUC-MSCs by isolating from human umbilical cord.