犬骨髓基质干细胞向成骨细胞诱导分化及细胞片层的制备  

Study of induced differentiation of bone marrow stromal stem cells into osteoblasts and preparation of cell sheets in dogs

作　　者：卜令学[1] 高振华[1] 荆恒[1] 陈立强[1] 刘凤芝[1] 李宁毅[1] 

机构地区：[1]青岛大学医学院附属医院口腔颌面外科,266003

出　　处：《现代口腔医学杂志》2010年第5期350-354,共5页Journal of Modern Stomatology

基　　金：国家自然科学基金资助项目(30872896);山东省自然科学基金资助项目(Y2008C77)

摘　　要：目的分离培养犬骨髓基质干细胞(bone marrow stromal cells,BMSCs)并向成骨细胞诱导分化,制备骨髓基质干细胞的细胞片层。方法密度梯度离心法分离犬骨髓基质干细胞,接种于DMEM成骨诱导培养基,向成骨细胞诱导分化,利用温度反应性培养皿的温度感应性,制备细胞片层。观察犬BMSCs、成骨细胞诱导分化及细胞片层形态学特点与生物学特性。结果分离培养出犬BMSCs,成功地向成骨细胞实现诱导分化,并观察到钙结节。利用温度反应性培养皿的温度感应性,收获了完整的细胞片层。结论细胞片层技术与传统的骨组织工程方法相结合,将有望构建出含板层骨结构的大块组织工程骨。Objective To isolate,culture and induce dog bone marrow stromal cells into osteoblasts in vitro,and to prepare cell sheets with induce-differentiated bone marrow stromal cells.Methods Bone marrow stromal cells were seeded in DMEM medium after isolated by density gradient centrifugation,and induced differentiation into osteoblasts,temperature-responsive culture dish was used to prepare cell sheets.Morphological features and biological properties of osteoblasts and cell sheets were observed by assay of methyl thiazolyl tetrazolium（MTT）,in-verted phase contrast microscope,alkaline phosphorase（ALP） stain and stain of calcified nodules.Results Dog bone marrow stromal cells were isolated and cultured,and were successfully induced into osteoblasts;calcium nodules was also observed.Complete cell sheets were harvested with temperature-responsive culture dishes.Conclusion It is expected to build larger tissue-engineered bone which has cortical bone structure by traditional methods of bone tissue engineering combing with cell-sheet engineering approach.

关 键 词：骨髓基质干细胞 细胞片层 组织工程骨 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]