原子力显微镜观察人脐带间充质干细胞:生物学特性与超微结构的关系  

Human umbilical cord mesenchymal stem cells under atomic force microscope:Correlation between biological characteristics and ultrastructure

作　　者：陈丽 吴本清 朱华民[2] 

机构地区：[1]暨南大学第二临床医学院,深圳市人民医院新生儿科,广东省深圳市518020 [2]暨南大学医学院,广东省广州市510632

出　　处：《中国组织工程研究与临床康复》2010年第6期996-1001,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：2009年度深圳市医学重点学科专项基金(20091998A03)~~

摘　　要：背景:细胞形态结构和功能密不可分,但目前对人脐带间充质干细胞超微结构的报道还很少。目的:探讨人脐带间充质干细胞的生物学特性与原子力显微镜下超微结构的关系。方法:采用酶消化法体外分离培养并扩增人脐带间充质干细胞,取第3代细胞,用原子力显微镜在接触模式下观察成像,通过流式细胞仪检测细胞免疫表型和细胞周期,油红O染色及碱性磷酸酶染色鉴定其成脂、成骨分化潜能。结果与结论:第3代人脐带间充质干细胞强表达CD44,CD29,低表达CD106,不表达CD34;有80%以上的细胞处在G0/G1期,增殖指数为19.9%;成脂诱导后,油红O染色可见胞浆中有大量桔红色的小脂滴;成骨诱导后,碱性磷酸酶染色可见立方形、多边形细胞的胞质染成棕褐色。原子力显微镜下人脐带间充质干细胞以长梭形为主,细胞骨架丝明显,并形成网状连接,这与其强大的增殖、迁移、分化功能相适应。BACKGROUND：The relationship between cellular morphosis and function is undetachable,but there is little investigation about ultrastructure of human umbilical cord mesenchymal stem cells （hUCMSCs）.OBJECTIVE：To study the relationship between the functions of hUCMSCs and their ultrastructure obtained by atomic force microscope （AFM）.METHODS：hUCMSCs were isolated,cultured,expanded after enzyme digestion.P3 cells were observed under AFM.Immunophenotype and cell cycle were analyzed by flow cytometry,as well as induction of the adipogenic,osteogenic differentiation of hUCMSCs were identified using Oil red O staining and alkaline phosphatase staining.RESULTS AND CONCLUSION：hUCMSCs at passage 3 were strongly positive for CD44 and CD29,weakly positive for CD106,but negative for hematopoietic marker CD34.Cells in G0/G1 phase accounted for 80%.Proliferation index was 19.9%.Following adipogenic induction,alkaline phosphatase staining demonstrated brown cytoplasm in cube and polygonal cells.AFM showed hUCMSCs were spindle shape,obvious cytoskeletal filament that connected into nets,which fit for the strong capacities for proliferation,migration and differentiation.

关 键 词：原子力显微镜 功能 超微结构 人脐带间充质干细胞 干细胞 

分 类 号：R394.2[医药卫生—医学遗传学]