骨髓间充质干细胞在特定微环境中自然分组向成骨细胞和内皮细胞分别分化的可行性研究  

Study of inducing bone mesenchymal stem cell to form osteoblast and endothelial cell in one microenvironment

作　　者：王嫣[1] 陈小菊[1] 徐杰[1] 于海生[1] 罗月苹[1] 周兰[2] 陈文直[3] 

机构地区：[1]重庆医科大学生物医学工程系省部共建超声医学工程国家重点实验室超声医学工程重庆市市级重点实验室,重庆400016 [2]重庆医科大学检验系,重庆400016 [3]重庆医科大学附属第二医院海扶肿瘤治疗中心,重庆400010

出　　处：《激光杂志》2011年第2期51-53,共3页Laser Journal

基　　金：国家重点基础研究发展计划-973计划项目(2011CB707902);国家自然科学基金青年科学基金项目(31000435);重庆市科委自然科学基金项目(CSTC;2009BB5261)

摘　　要：目的:观察骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)在特定的微环境中是否具有自然分组向成骨细胞和内皮细胞(endothelial cell,EC)两种不同细胞同时分化的可能。方法:将BMSCs采用成骨诱导剂和成内皮细胞诱导剂以1:1的比例混合培养12天。倒置显微镜和HE染色观察细胞的形态变化;进行骨钙素和vWF免疫细胞化学染色,以探讨BMSCs向成骨细胞和EC的分化情况;用图像分析系统测定BMSCs向EC的分化率;激光扫描共聚焦显微镜(lasers canning confocal microscope,LSCM)进一步观察细胞向两个方向分化的情况。结果:使用倒置显微镜和HE染色可在一个视野里见到形态不同的几种细胞。分别进行骨钙素和vWF免疫细胞化学染色均为阳性,体视分析EC面密度为14.1%。LSCM显示细胞同时表达两种抗原。结论:在由成骨诱导剂和成内皮细胞诱导剂混合而成的特定微环境中,BMSCs可能具有被诱导后自然分组向两个方向同时分化的潜能。Objective：To explore the possibility of inducing BMSCs to differentiate into osteoblasts and EC in same microenvironment simultaneously.Method： BMSCs were cultured in the mixing medium which contains dexamethasone,ascorbic acid,β-glycerophosphate,VEGF and L-glutamine for 12 days.Cells were observed by microscope and HE stain,incubated with anti-vWF and anti-Osteocalcin,and analysed the differentiation rates from BMSCs to EC by image analysis.Immunofluorescence with anti-vWF and anti-Osteocalcin were observed by laser scanning confocal microscopy（LSCM）.Results： The different morphous can be observed by light microscope and HE stain： some cells formed calcified nodules and some cells formed a tube similarity.Osteocalcin and vWF were positive in the two kinds of cells;the density of EC was 14.1%.After fluorescence immunocytochemistry staining, two colors can be detected in cells simultaneously with LSCM.The mix of two colors was yellow,but the colors of some cells were much red,some were much green.Conclusion： BMSCs could differentiate into osteoblasts and EC in the same microenvironment when precisely induced.

关 键 词：组织工程 干细胞 分化 

分 类 号：R318[医药卫生—生物医学工程]