【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。更多还原

【Abstract】 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 chain α-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 of α-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.更多还原