| Objectives To establish a method of isolating and culturing human bone marrow-derived mesenchymal stem cells(BMSCs) ; to investigate the feasibility of collagen I as a kind of scaffolds using in cartilage tissue engineering ; to discuss the best cellular density and culture time in cartilage tissue engineering which use BMSCs as seeding cell and collagen I as scaffolds .Methods A volume of 6 ml bone marrow was collected from six volunteer donors respectively. BMSCs were cultured in the Dulbecco's Modified Eagle's Medium (DMEM) with low glucose containing 10% fetal bovine . Depending their different cellular density combined with collagen I ,BMSCs from the third passage were divided into four groups :A 1×104/ml; B 1×105/ml; C 1×106/ml; D 5×106/ml. Then, all groups were inductive by the DMEM which has 10ng/ml TGF-β1 Thegrowth of the cultured BMSCs was observed.Imrnunohistochemistry and in situ hybridization were applied to detect the expression of collagen type II .The synthetic proteoglycans were detected by toluidine blue staining.Results We can get BMSCs through this method. After two weeks inductive culturing The cells of the B,C,D groups were positively stained by toluidine blue, immunohistochemistry and in situ hybridization analysis proved strong type II collagen expression. And their GAG content were sharply increased from the second week to fourweek(4week>3week>2week). But their GAG content were stop increasing in the fifths week.
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