基于骨髓间充质干细胞体外构建组织工程心肌样组织的实验研究  

The Experimental Study on Constructing the Tissue Engineered Myocardium-like Tissue in vitro with Bone Mesenchymal Stem Cells

作　　者：邢万红[1] 马捷[1] 郭光伟[1] 李新华[1] 席虎玉[1] 

机构地区：[1]山西医科大学第二医院,山西太原030001

出　　处：《透析与人工器官》2012年第4期8-13,共6页Chinese Journal of Dialysis and Artificial Organs

基　　金：山西省科技攻关项目资助(NO.20080311061-2)

摘　　要：目的本文旨在探讨应用婴幼儿骨髓间充质干细胞(bone mesenchymal stem cells,BMSCs)和PLGA体外构建三维心肌样组织。方法分离培养CHD婴幼儿胸骨BMSCs,经5-氮杂胞(5-Azacytidine,5-Aza)苷诱导分化为心肌细胞,种植到可降解的载体-聚乳酸/羟基乙酸的共聚物(polyactic acid/glycolic acid homoconjugate,PLGA)上,加入含有10%胎牛血清的LG-DMEM培养液,摇床培养1周后种植到裸鼠背部皮下。实验终止后通过组织学、免疫化学和Real-timePCR等对构建的组织工程化心肌样组织进行评价。结果婴幼儿BMSCs具有稳定生长和快速增殖的特性。免疫组织化学染色结果表明,体外构建的组织工程化心肌α-actin,desmin,Cx-43和cTnI等抗体染色阳性。透射电镜显示,构建的组织工程花样心肌组织具有肌丝,缝隙连接和闰盘样结构形成。Real-timePCR等结果表明,构建的工程化心肌样组织具有自体心肌的特性。结论以婴幼儿BMSCs为种子细胞,PLGA为支架能够体外构建工程化心肌样组织。本文的研究结果为体外研制用来修复心肌缺损或扩大先天性心脏病流出道如室间隔缺损、法罗氏四联症等工程化心肌修复材料奠定了基础。Objective This paper aims to constructing in vitro engineering 3-D myocardium-like tissue with bone mesenchymal stem cells(BMSCs)of infants and poly-lactic-co-glycolic acid(PLGA).%Methods Bone marrow was obtained from the sternal marrow cavum outflow of infant with congenital heart disease(CHD)undergoing cardiac surgery.BMSCs were obtained by density gradient centrifugation.The cell-scaffold complex has been cultured in the shake cultivation for 1 week,then the complex has been planted in the dorse of the nude mouse.When the experiment had been finished,the histology,immunology,real time PCR and so on were done.%Results The BMSCs of infant with congenital heart disease have the properties of the stable growth and the rapid proliferation.The immunohistochemistry showed that tissue engineered myocardium constructed in vitro expressed some cardiac related proteins such as α-actin,Cx-43,desmine,cTNI and so on.The transparent myofilaments,gap junctions and intercalated disk-like structure formation could be observed in the 3D tissue-like constructs by transmission electron microscope(TEM).The engineered myocardium-like tissue had the auto-myocardial property as assessed by real-time PCR and so on.Conclusion The engineered myocardial tissue-like constructs could be built with infant BMSCs and PLGA.Our results may provide a foundation for the long engineering myocardial material in repairing the defect or augmenting the tract in CHD,such as ventricular septal defect,tetralogy of Fallot and so on.

关 键 词：组织工程 骨髓间充质干细胞 5-氮杂胞甘 心肌 体外 

分 类 号：R318.01[医药卫生—生物医学工程]