碱性成纤维细胞生长因子联合间充质干细胞构建组织工程瓣膜的研究  

Construction of tissue engineered heart valve using basic fubribkast growth factir and mesenchymal stem cells

作　　者：洪昊[1] 苏伟[1] 董念国[1] 史嘉玮[1] 

机构地区：[1]华中科技大学协和医院心外科华中科技大学同济医学院心血管病研究所,武汉430022

出　　处：《临床心血管病杂志》2009年第11期849-852,共4页Journal of Clinical Cardiology

基　　金：国家自然科学基金资助项目(No:30600608和30872540);国家高技术研究发展计划资助项目(2009AA03Z420)

摘　　要：目的:探讨采用碱性成纤维细胞生长因子(bFGF)联合间充质干细胞(MSCs)体外构建组织工程瓣膜(TEHV)过程中,MSCs表型特性和基质金属蛋白酶及其抑制物的表达情况。方法:分离培养大鼠MSCs,进行细胞鉴定后种植于去细胞瓣叶支架上。将瓣叶置于含5ng/mlbFGF的培养液中,培养14d构建的TEHV作为A组;B组除培养液中不添加bFGF外,其余同A组;C组为大鼠肌成纤维细胞构建TEHV的对照组。RT-PCR检测各组TEHV中α-SMA、MMP-13和TIMP-1mRNA的表达。结果:MSCs表达CD29(94.82%)和CD44(93.59%)。A组与C组α-SMA、MMP-13和TIMP-1mRNA表达比较均差异无统计学意义(P>0.05),但均高于B组(P<0.05)。结论:采用bFGF联合MSCs体外构建TEHV过程中,通过bFGF的调控作用,可以使MSCs的表型特性和基质金属蛋白酶及其抑制物表达与肌成纤维细胞相当。Objective：To study during the process of tissue engineered heart valve （TEHV） construction in vitro using basic fibroblast growth factor （bFGF） and mesenchymal stem cells （MSCs）, the phenotype of MSCs and its expression of matrix metalloproteinase and tissue inhibitor of metalloproteinase were investigated. Method： MSCs were isolated and cultured. After cell identification, MSCs were seeded onto decellularized aortic valve leaflet scaffold to constuet TEHV in each group. In group A, TEHV were cutlured with DMEM contained 5 ng/ml bFGF in vitro for 14 d. In group B, TEHV were cutlured with DMEM only for 14 d. In control group C, TEHV were constructed using myofibroblasts as seed cells. The expression of α-SMA, MMP-13 and TIMP-1 mRNA were analyzed by reverse transcription-polymerase chain reaction （RT-PCR）. Result：MSCs expressed CD29 （94. 82%） and CD44 （93.59%）. Group A and control group C showed comparable expression of α-SMA, MMP-13 and TIMP-1 mRNA （P〉0.05）. However the mRNA expression were significantly inceased in group A and control group C, compared with lower values of group B （P〈0. 05）. Conclusion： During the process of TEHV construction in vitro, when regulated by bFGF, the phenotype of MSCs was the same as myofibroblasts and its expression of matrix metalloproteinase and tissue inhibitor of metalloproteinase was equal to myofibroblasts.

关 键 词：间充质干细胞 碱性成纤维细胞生长因子 组织工程瓣膜 基质金属蛋白酶 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]