应用细胞片层技术构建组织工程骨  

Construction of tissue-engineered bone using cell sheet technology

作　　者：陈涛[1] 王艳辉[1] 卜令学[1] 荆恒[1] 高振华[1] 谭帅[1] 王玲玲[1] 李宁毅[2] 

机构地区：[1]青岛大学医学院附属医院医学影像科,山东省青岛市266003 [2]青岛大学医学院口腔颌面外科,山东省青岛市266003

出　　处：《中国组织工程研究与临床康复》2011年第2期191-195,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

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);课题名称:应用细胞片层技术构建血管化组织工程骨的实验研究~~

摘　　要：背景:传统的骨组织工程学在收获和转移细胞方面仍存在诸多不足,细胞片层技术是收获种子细胞和对种子细胞进行转移的一项新技术。目的:应用细胞片层技术与传统骨组织工程相结合的方法构建组织工程骨。方法:密度梯度离心法分离培养犬骨髓间充质干细胞,诱导其向成骨细胞分化。将诱导后的骨髓间充质干细胞接种至温度反应性培养皿中,先37℃饱和湿度培养,然后降温至20℃制备细胞片层;将犬脱钙骨基质/富血小板血浆/骨髓间充质干细胞细胞片层/骨髓间充质干细胞复合体植入犬左侧背阔肌下,术后4,8,12周,观察成骨情况。结果与结论:当温度降至20℃时,骨髓间充质干细胞从温度反应性培养皿上完全脱落,形成细胞片层,将其覆盖于犬脱钙骨基质/富血小板血浆/骨髓间充质干细胞后植入犬背阔肌下,其成骨效果明显好于不加细胞片层对照侧。说明应用细胞片层技术与传统的骨组织工程方法相结合可构建出较理想的组织工程骨。BACKGROUND：Traditional bone tissue engineering faces shortcomings in harvesting or transferring cells.Cell sheet technology is a new skill for cell harvesting or transferring.OBJECTIVE：To construct tissue-engineered bone using cell sheet technology and method of traditional bone tissue engineering.METHODS：Canine bone marrow mesenchymal stem cells （BMSCs） were isolated with the method of density gradient centrifugation and cultured.BMSCs were induced to differentiate into osteoblasts and cultured in temperature-responsive culture dishes at 37℃,5% CO2 saturated humidity.BMSCs cell sheet was prepared when temperature was changed to 20℃;demineralized bone matrix and platelet rich plasma （PRP） were prepared.The complex of DBM/PRP/BMSCs cell sheet/BMSCs was construsted and implanted under the left latissimus dorsi muscle.The condition of osteogenesis was observed at 4,8 and 12 weeks after operation.RESULTS AND CONCLUSION：When temperature dropped at 20℃,BMSCs detached automatically from the temperature-responsive culture dishes and formed an intact cell sheet.The osteogenesis of the DBM/PRP/BMSCs cell sheet/BMSCs group was better than that of the DBM/PRP/BMSCs group.Cell sheet technology combined with traditional bone tissue provides a new way for construction of functional tissue-engineered bone.

关 键 词：细胞片层 骨髓间充质干细胞 温度反应性培养皿 成骨细胞 骨组织工程 

分 类 号：R318[医药卫生—生物医学工程]