Transwell共培养条件下诱导脂肪干细胞成骨能力的改变  

Osteogenic efficiency of induced adipose-derived stem cells under Transwell co-cultured condition

作　　者：孙仕晨 董腾哲 黄昕[1] 张云龙[1] 陈刚[1] 

机构地区：[1]天津医科大学口腔医院口腔颌面外科,天津市300070

出　　处：《中国组织工程研究》2016年第28期4155-4161,共7页Chinese Journal of Tissue Engineering Research

基　　金：教育部留学回国人员科研启动基金资助项目[教外司留(2012)940]~~

摘　　要：背景:在共培养条件下,间充质干细胞可调节成骨细胞的分化和成骨能力。目的:研究成骨细胞前体与未分化的骨髓间充质干细胞、脐带间充质干细胞和胎盘间充质干细胞在矿化培养液中共培养对其成骨效能的影响。方法:脂肪干细胞成骨诱导7 d后获得诱导后的脂肪干细胞,将其分别与上述骨髓、其脐带和胎盘来源的间充质干细胞在Transwell小室中进行间接共培养。实验组根据间充质干细胞种类不同将其分为骨髓来源组、脐带来源组和胎盘来源组,对照组为诱导后的脂肪干细胞单独培养。在不同实验间期通过检测碱性磷酸酶活性以及定量分析钙基质沉积情况,观察不同组织来源的间充质干细胞对诱导后脂肪干细胞成骨效能的影响。结果与结论:(1)碱性磷酸酶表达活性:间接共培养条件下实验组各组碱性磷酸酶活性显著高于对照组(P<0.05),实验组中骨髓来源组碱性磷酸酶活性高于脐带来源组和胎盘来源组(P<0.05);(2)钙化基质定量分析:实验组各组均显著高于对照组(P<0.05),实验组中脐带来源组高于骨髓来源组和胎盘来源组(P<0.05);(3)结果表明:诱导后的脂肪干细胞在间接共培养条件下受间充质干细胞的调节成骨能力明显提高。BACKGROUND：Under co-culture conditions, mesenchymal stem cel s could regulate osteogenic differentiation and osteogenesis of osteoblasts. OBJECTIVE：To observe the osteogenic efficiency of osteoblastic precursor cel s co-cultured with undifferentiated bone marrow-derived mesenchymal stem cel s, umbilical cord-derived mesenchymal stem cel s, or placenta-derived mesenchymal stem cel s in mineralization medium. METHODS：Adipose-derived stem cel s were induced in osteogenic differentiation medium for 7 days before being indirectly co-cultured with undifferentiated mesenchymal stem cel s isolated from different tissues （bone marrow group, umbilical cord group and placenta group） in Transwel plates. Induced adipose-derived stem cel s cultured alone served as control group. At different experimental intervals, quantitative analysis of alkaline phosphatase activity and calcified matrix was preformed to observe the effects of mesenchymal stem cel s from different sources on the osteogenic efficiency of induced adipose-derived stem cel s. RESULTS AND CONCLUSION：Expression of alkaline phosphatase was significantly higher in different experimental groups than the control group （P〈0.05）, and it was also higher in the bone marrow group than the umbilical cord and placenta groups （P〈0.05）. Quantitative analysis of calcified matrix revealed that the experimental groups were significantly higher than the control group （P〈0.05）;and in experimental groups, the umbilical cord group was higher than bone marrow group and placenta group （P〈0.05）. These findings indicate that the osteogenic efficiency of induced adipose-derived stem cel s is improved dramatical y under co-culture conditions.

关 键 词：干细胞 组织工程 碱性磷酸酶 脂肪干细胞 骨髓间充质干细胞 脐带间充质干细胞 胎盘间充质干细胞成骨 Transwell共培养 成骨细胞前体 分化 微环境 基质 诱导 

分 类 号：R394.2[医药卫生—医学遗传学]