中国冶金工业医学杂志

基因转染骨髓基质干细胞构建组织工程心脏瓣膜的实验研究

作者：徐瑞剑王志强

分类号：中图分类号：R 605.974

出版年,卷(期):页码：2015,32(1):1-4

摘要：

目的探索一种行之有效的方法增强细胞黏附力，加快组织工程心脏瓣膜的体外构建。方法通过纤维连接蛋白羧基端细胞结合域(FnCBD64)的原核表达、提取和纯化,构建FnCBD64基因重组腺病毒载体Ad.FnCBD64,并进行绵羊骨髓基质干细胞（BMSCs）的体外分离培养及生物学特性分析,制备去细胞猪主动脉瓣,在无菌条件下将脱细胞瓣叶圆片置入96孔板中(心室面向上)，分别给予Fn、FnCTD64牛血清白蛋白（BSA）预先包埋，未预包埋作为对照组。细胞悬液接种，每只瓣叶种植细胞数为4×105个。在开始种植细胞的第8天取瓣叶进行细胞计数，电镜扫描MTT检测。结果对照组在经过脉动冲刷实验高流量液体的冲刷以后，其表面的骨髓间质干细胞几近完全脱落，电镜扫描下为裸露的瓣膜支架；而预涂FnCBD64组虽然经过高流量的流体冲刷，有稀疏的细胞残留，细胞间有瓣膜支架的结构；转染Ad.FnCBD64组瓣膜表面存留的细胞比预涂FnCBD64组明显多，某些部位细胞呈复层生长，细胞可见排列极性改变，有向活体内细胞转化的趋势。结论预涂FnCBD64和转染Ad.FnCBD64均能明显增强BMSCs在去细胞猪主动脉瓣叶及整体瓣膜上的黏附，而转染Ad.FnCBD64的效果更好。关键词：基因转染；骨髓基质干细胞；组织工程；心脏瓣膜

ObjectiveTo explore an effective way to enhance cell adhesion to accelerate in vitro tissue engineering heart valve constructed.MethodsTo construct recombinant adenovirus vector Ad.FnCBD64 by fibronectin cell binding domain(FnCBD64)prokaryotic expression,extraction,sheep marrow stromal cells(BMSCs)in vitro culture and biological characteristics analysis,preparation acellular porcine aortic valve,acellular leaflets wafer under sterile conditions into 96\|well plates(ventricular side up),Fn,FnCTD64,bovine serum albumin BSA entrapped in advance and not pre\|embedded as a control group.Cell suspension was inoculated,number of cells each leaflet was 4×l05 which were counted on the eighth day from the beginning of cells growing,MTT measured with electron microscopy.ResultsIn the control group,marrow mesenchymal stem cells on the surface almost completely were off and appeared bare valve stents under electron microscope after scouring by pulsating fluid with high flow bone;while in precoated FnCBD64 group,even after high\|flow fluid erosion,there were sparse cells remained and valve stents among cells;cells retained on valve surface in Ad.FnCBD64 group were significantly more than that in the pre\|coat FnCBD64 group.Some cells grown stratified,cell polarity changed and trended to transformation of in vivo cell.ConclusionBoth pre\|coated FnCBD64 and transfection Ad.FnCBD64 could significantly enhance the adhesion of BMSCs on acellular porcine aortic valve leaflets and the whole leaflets,while transfection Ad.FnCBD64 is better.Key Words:Gene transfection;Bone marrow stromal cells;Tissue engineering;Heart valve

基金项目：

内蒙古自治区自然科学基金项目（项目编号2009MS1148）

作者简介：

参考文献：

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