犬同种异体脱钙骨基质复合骨髓基质干细胞片层与内皮祖细胞的体外培养

Study of bone marrow stroma stem cells cell-sheet and endothelial progenitor cells cultured in vitro on allogenic demineralized bone matrix in mongrel

目的:观察犬骨髓基质干细胞(bone marrow stroma stem cells,BMSCs)细胞片层及内皮祖细胞(endothelial progenitor cells,EPCs)在脱钙骨基质(demineralized bone matrix,DBM)上的生长情况,探讨利用其复合体构建功能性血管化组织工程骨的可行性。方法:应用细胞片层技术制备BMSCs细胞片层,培养犬内皮祖细胞,制备犬同种异体DBM。将DBM分别与BMSCs细胞片层、内皮祖细胞复合。测量DBM孔隙率,倒置相差显微镜、扫描电镜下观察DBM的结构及BMSCs细胞片层、EPCs在DBM上的附着、生长情况。结果:成功构建DBM/BMSCs细胞片层、DBM/EPCs复合体,BMSCs细胞片层、EPCs在DBM上的附着、生长状况良好,增殖迅速。结论:DBM/BMSCs细胞片层、DBM/EPCs复合体能够为生成具有骨样结构的组织工程骨提供良好的条件,并为组织工程骨的血管化创造有利的环境。

PURPOSE： To investigate the growth situation of bone marrow stroma stem cells（BMSCs） cell-sheet and endothelial progenitor cells（EPCs） on demineralized bone matrix（DBM） and explore the effect of DBM/BMSCs cell-sheet and DBM/EPCs complex on the construction of functional and vascularized tissue-engineered bone in mongrel.METHODS： BMSC cell-sheet was prepared with cell-sheet engineering approach,EPCs were isolated from canine bone marrow and DBM was prepared from homologous bone.BMSCs cell-sheet and EPCs were inoculated on DBM respectively and observed by inverted phase contrast microscope and scanning electron microscope.The porosity of DBM was measured.RESULTS： The DBM/BMSCs cell-sheet and DBM/EPCs complex were successfully constructed.The BMSCs cell-sheet and EPCs adhered to DBM well and grew rapidly.CONCLUSIONS： The BMSCs cell-sheet and EPCs have good biocompatibility with DBM.The complex of DBM/ BMSCs cell-sheet,DBM/EPCs could provide good conditions to acquire functional and vascularized tissue-engineered bone.