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Background:Cartilage defects has become a major problem for treatment of femoral head collapse in late stage of osteonecrosis of femoral head(ONFH).It was new theory that combined culture of bone marrow stromal stem cells in vitro and Bio-gide collagen membrane to repair cartilage defects.Objective:This study was to study canine bone marrow stromal stem cells isolation,culture and identification methods of ONFH as a cartilage defect repair tissue engineering seed cells is feasible.Methods:In vitro cultured BMSCs,the morphology,cell surface antigen,induced chondrocyte differentiation and identification; will BMSCs with Bio-gide membrane collagen in vitro co-culture by inverted phase contrast microscope,scanning electron microscopy observation.Results:The combination of bone marrow adherent blood centrifugation screening method to get high purity active BMSCs.BMSCs in the Bio-gide membrane attached to collagen proliferation,and multi-layer growth.Conclusion:BMSCs easily obtained and expanded differentiation ability.BMSCs can be used as ONFH in tissue engineering of cartilage defects source of seed cells.Bio-gide membrane collagen as a carrier of cell growth.