SD大鼠乳鼠脱细胞心脏支架的细胞相容性评价  

Cell biocompatibility of an acellular heart scaffold of Sprague-Dawley neonatal rats

作　　者：韦雅淑 徐亦辰 赵文婧[1] 王慧丰[1] 刘红静 谢丹尼[1,2] 陈维平 Wei Ya-shu;Xu Yi-chen;Zhao Wen-jing;Wang Hui-feng;Liu Hong-jing;Xie Dan-ni;Chen Wei-ping(Department of Histology and Embryology,Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China;Guangxi Zhuang Autonomous Region Family Planning Research Center,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学组织学与胚胎学教研室,广西壮族自治区南宁市530021 [2]广西壮族自治区计划生育研究中心,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2018年第2期261-266,共6页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金(2012GXNSFAA053126);广西科技厅资助项目(桂科自0832157;桂科攻0592007-1G);广西急诊与医学救援人才小高地.广西高校急诊医学重点实验室开放课题(GXJZ201505);广西医科大学青年基金(GXMUYSF201528);2017年度广西高校中青年教师基础能力提升项目(2017KY0096);广西再生医学重点实验室开放课题资助项目(桂再重开201703);广西壮族自治区人口和计划生育委员会科研基金(桂人口计生研1108)~~

摘　　要：背景:脱细胞心脏支架材料具有低致敏性和天然的三维结构、具有良好的生物相容性和细胞亲和性等优点,成为心肌组织工程研究的热点。目的:探究SD大鼠乳鼠脱细胞心脏支架的细胞相容性。方法:选择第3代SD大鼠乳鼠骨髓间充质干细胞与SD大鼠乳鼠脱细胞心脏支架共培养7,14 d,构建细胞-支架复合物,采用苏木精-伊红染色、扫描电镜观察骨髓间充质干细胞在支架内的生长情况。采用心肌组织裂解液对细胞-支架复合物进行定向分化诱导,连续培养14 d,以平面定向分化诱导连续培养14,20 d的骨髓间充质干细胞为对照组,采用RT-PCR方法检测骨髓间充质干细胞内肌球蛋白重链α-MHC、锌指转录因子GATA-4的表达。结果与结论:(1)苏木精-伊红染色显示,细胞-支架复合物中存在大量嗜碱性细胞核,细胞-支架复合物共培养14 d的细胞数量明显多于共培养7 d;扫描电镜下观察显示,共培养14 d的细胞-支架复合物纤维表面有大量细胞黏附;(2)平面定向分化诱导连续培养14,20 d的骨髓间充质干细胞分别呈竹节样、肌管样结构。在定向分化诱导连续培养14 d的细胞-支架复合物中可检测到α-MHC、GATA-4的表达,α-MHC、GATA-4的表达量分别与竹节样细胞和肌管样结构阶段相当;(3)结果表明脱细胞心脏支架的细胞相容性良好。BACKGROUND:Owing to the advantages of low sensitization and natural three-dimensional structure,good biocompatibility and cell affinity,acellular heart scaffold materials are of great current interest in cardiac tissue engineering.OBJECTIVE:To investigate the cytocompatibility of an acellular heart scaffold of neonatal rats.METHODS:In order to construct the seed cell-scaffold complex,passage 3 bone marrow mesenchymal stem cells(BMSCs)of Sprague-Dawley neonatal rats were cultured with an acellular heart scaffold of Sprague-Dawley neonatal rats for 7 and 14 days.Hematoxylin eosin staining and scanning electron microscopy were used to observe the growth of BMSCs in the scaffold.The cell-scaffold complex was induced in myocardial tissue lysate for 14 days.BMSCs with planar orientation differentiation for 14 and 20 days were used as control group.RT-PCR was used to detect the expression of myosin heavy chainα-MHC and zinc finger transcription factor GATA-4 in BMSCs.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining showed the acellular heart scaffold contained a large number of eosinophilic fibrous structures,and the cell number of cell-scaffold complex after co-culture for 14 days was higher than that after co-culture for 7 days.Under the scanning electron microscope,a large amount of cells adhered to the fiber surface of the acellular scaffold at 14 days of co-culture.(2)BMSCs with planar orientation differentiation for 14 and 20 days had the bamboo-like and myotube-like structures.In the cell-scaffold complex with planar orientation differentiation for 14 days,the expression ofα-MHC and GATA-4 could be detected,and their expression levels fulfilled the requirement for the presence of bamboo-like cells and myotube-like structure.These results indicate that the acellular heart scaffold exhibits good cytocompatibility.

关 键 词：干细胞 肌细胞 心脏 肌球蛋白重链 组织工程 

分 类 号：R318[医药卫生—生物医学工程]