SPIO标记骨髓间充质干细胞和软骨细胞共培养的研究  

Research of co-culture of chondrocyts and bone marrow mesenchymal stem cells labeled by SPIO in vitro

作　　者：詹兴旺 姜艳[2] 王文良[3] 

机构地区：[1]武警8750部队医院,云南蒙自661100 [2]红河学院理学院,云南蒙自661100 [3]武警医学院附属医院骨科,天津300162

出　　处：《中国矫形外科杂志》2011年第9期769-773,共5页Orthopedic Journal of China

基　　金：国家自然科学基金项目(编号:30740089);天津市自然科学基金(编号:10JCYBJC10800);武警医学院科研基金项目(编号:WZK200902)

摘　　要：[目的]探讨SPIO标记骨髓间充质干细胞和软骨细胞共培养的可行性,为临床修复关节软骨损伤寻找新途径。[方法]分离、扩增新西兰大白兔骨髓间充质干细胞(BMSCs)和软骨细胞,根据SPIO标记(50μg/m l)和不同培养环境(共培养、单独培养)分4组,每天在倒置显微镜观察共培养后BMSCs的形态变化,共培养14 d后,免疫组化检测Ⅱ型胶原表达情况,阿利新蓝法检测蛋白多糖(GAG)表达水平。[结果]共培养7 d后标记的部分BM-SCs变圆,14 d时BMSCs形态高度分化与成熟软骨细胞相似,其蛋白多糖和Ⅱ型胶原的基因转录和蛋白表达均增高,明显优于对照组,差异具有显著性意义(P<0.01)。[结论]1.软骨细胞微环境能有效诱导BMSCs向软骨细胞分化;2.SPIO可以安全、有效地标记BMSCs。[Objective]To investigate the feasibility of co-culture of bone marrow mesenchymal stem cells（BMSCs） labeled by SPIO and chondrocytes so as to provide a new clinical approach for repairing damaged articular cartilage.[Methods]BMSCs and chondrocytes were exracted from adult New Zealand white rabbits.These cells were isolated and cultured in different environments（co-culture,single-culture）.The shapes of BMSCs before and after co-culture were observed daily by inverted microscopy.The expression of aggrecan and type II collagen in different groups were detected with alcian blue staining and immunohistochemistry at 14 days after co-culture in vitro.[Results]The shapes of BMSCs became round at 7 days after co-culture in vitro.At 14 days after co-culture with chondrocytes,typeⅡcollagen and aggrecan were markedly increased.There were significant differences between co-culture group and the other single-culture groups（P0.01）.[Conclusion]1.Chondrocytes may provide chondrogenic microenvironment to induce chondrogenic differentiation of BMSCs in vitro efficiently.2.BMSCs can be marked by SPIO safely and efficiently.

关 键 词：SPIO 骨髓间充质干细胞 软骨细胞 共培养 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]