绿色荧光蛋白标记大鼠骨骼肌卫星细胞与聚羟基乙酸的生物相容性  

Biocompatibility of labeled green fluorescent protein skeletal muscle satellite cells grown on polygly-colic acid fibers

作　　者：王营 王曦龙 章俊 王强 施国伟 Wang Ying;Wang Xilong;Zhang Jun;Wang Qiang;Shi Guowei(Department of Urology,the Fifth People＇s Hospital of Shanghai,Fudan University,Shanghai 200240,China)

机构地区：[1]复旦大学附属上海市第五人民医院泌尿外科,上海200240

出　　处：《中华实验外科杂志》2018年第11期2014-2016,共3页Chinese Journal of Experimental Surgery

基　　金：上海市医学重点专科基金（ZK2015804）;上海市卫生和计划生育委员会科研课题青年项目（20164Y0059）;闵行区科委自然科学基金（2016MHZ37）

摘　　要：目的建立大鼠骨骼肌卫星细胞的分离和培养方法，探讨骨骼肌卫星细胞复合聚羟基乙酸（PGA）材料在体外共培养的可行性。方法酶消化法获取原代骨骼肌卫星细胞，经体外培养和扩增后，组织学检测骨骼肌卫星细胞标志性蛋白的表达，将慢病毒载体介导绿色荧光蛋白（GFP）转染后的骨骼肌卫星细胞接种于PGA上，形成骨骼肌卫星细胞-PGA材料复合物，体外培养1周后，进行光镜及扫描电镜观察。结果培养的骨骼肌卫星细胞表达结蛋白（Desmin）和波形蛋白（Vimentin）阳性，光镜下观察显示转染后的骨骼肌卫星细胞在PGA纤维上增殖并分泌大量的细胞外基质，扫描电镜显示细胞分泌的细胞外基质将PGA纤维之间的孔隙充满。结论转染后的骨骼肌卫星细胞复合PGA后生长良好，GFP可作为骨骼肌卫星细胞研究的示踪标记。Objective To establish a method of isolation and culture of the rat skeletal muscle satellite cells. To investigate the feasibility of in vitro constructing cell-polyglycolic acid （PGA） complex from skeletal muscle satellite cells seeded on PGA. Methods Primary skeletal muscle satellite cells were harvested by enzyme. After series culture and expansion in vitro, Immunohistochemistry staining was carried out to detect the immunophenotypes of skeletal muscle satellite cells. After skeletal muscle satellite cells were transfected by lentiviral vector carrying green fluorescent protein （GFP）, skeletal muscle cell- PGA complex was constructed by seeding transfected skeletal muscle satellite cells on PGA fibers. The cell- PGA complex was examined by light microscope and scanning electron microscope （ SEM ）. Results Immunohistochemistry staining showed that skeletal muscle satellite cells expressed Desmin and Vimentin. Microscopic observation showed the production of rich extracellular matrices by transfected skeletal muscle satellite cells was observed in the cell-scaffold construct, SEM observations showed that the secreted matrices filled the space between the PGA fibers. Conclusion Transfected skeletal muscle satellite cells have good biocompatibility with PGA. Imaging with expression of GFP facilitates the research on skeletal muscle satellite cells application in tissue engineering in vivo.

关 键 词：骨骼肌卫星细胞 绿色荧光蛋白 构建 组织工程 

分 类 号：R318.08[医药卫生—生物医学工程]