碱性成纤维细胞生长因子和骨髓间充质干细胞构建的组织工程心脏瓣膜赖氨酰氧化酶的表达  

Expression of lysyl oxidase in tissue engineering heart valve established by basic fibroblast growth factor and bone marrow mesenchymal stem cells

作　　者：洪昊[1] 苏伟[1] 董念国[1] 史嘉玮[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院心血管外科,武汉430022

出　　处：《中华生物医学工程杂志》2008年第6期400-404,共5页Chinese Journal of Biomedical Engineering

基　　金：国家自然科学基金（30571839、30600608）;国家高技术研究发展计划（2009AA03Z420）

摘　　要：目的探讨碱性成纤维细胞生长因子（bFGF）和骨髓间充质干细胞（MSC）构建组织工程心脏瓣膜（TEHV）及其赖氨酰氧化酶（LOX）的表达。方法贴壁培养法分离、培养和纯化大鼠MSC，取第3代种植于去细胞瓣叶支架上。分别将瓣叶置于含10μg／LbFGF的培养液（A组）、普通培养液（B组）中构建TEHV。大鼠肌成纤维细胞构建的TEHV为C组。培养14d后，采用Amplex red荧光法检测各组中的LOX蛋白含量，RT-PCR检测LOXmRNA表达。结果B组的LOX蛋白含量和mRNA表达[（0．137±0．003）mg／L，2．08±0．03]高于A组[（0．124±0．002）mg／L，0．87±0．01]和C组[（0．127±0．0HD2）mg／L，0．90＋0．01]（P〈0．05），A、C两组差异无统计学意义（P〉0．05）。结论bFGF可能通过5降低MSC的LOX表达，使MSC构建的TEHV达到类似肌成纤维细胞构建的TEHV。Objective To study tissue engineering heart valve （TEHV） established by basic fibroblast growth factor （bFGF） and bone marrow mesenchymal stem cells （MSCs） and expression of lysyt oxidase （LOX） in TEHV. Methods MSCs were isolated, cultured and purified by using adherent culture. MSCs of the third generation were seeded onto decellularized aortic valve leaflet scaffold. In group A, TEHVs were cutlured with DMEM media containing 10 μg/L bFGF. In group B, TEHVs were cutlured with DMEM media only. In group C, TEHVs were established by using myofibroblasts as seed cells. All the groups were cultured in vitro for 14 d. Level of LOX in each group was detected by Amplex red method and expression of LOX mRNA in each group was analyzed by reverse transcription-polymerase chain reaction. Results The level of LOX and expression of LOX mRNA [ （0.137±0.003 ） mg/L, 2.08±0.03 ] in group B were significantly higher than those in group A [ （0.124±0.002） mg/L,0.87±0.01 ] and group C [ （0.127±0.002） mg/L,0.90±0.01 ] （P〈0.05）. Group A and group C showed similar level of LOX and expression of LOX mRNA （P〉0.05）. Conclusion Expression of LOX in MSCs can be decreased by bFGF, so that TEHV established by MSCs can reach the same level as TEHV established by myofibroblasts.

关 键 词：碱性成纤维细胞生长因子 问质干细胞 组织工程 细胞培养 心脏瓣膜 赖氨酰氧化酶 

分 类 号：R318[医药卫生—生物医学工程]