大鼠骨髓间充质干细胞体外心肌定向诱导及心肌组织构建研究  

Study on induction of bone marrow mesenchymal stem cells into cardiomyocyte and construction of myocardial tissue in vitro

作　　者：王宁[1] 邢万红[1] 李新华[1] 陈平[1] 

机构地区：[1]山西医科大学第二医院心胸外科,太原030001

出　　处：《国际生物医学工程杂志》2010年第3期163-166,F0003,共5页International Journal of Biomedical Engineering

基　　金：山西省科技攻关项目（20080311061-2）;太原市科技发展项目（0904052）

摘　　要：目的探讨大鼠骨髓间充质干细胞（BMSCs）向心肌定向诱导分化及诱导后BMSCs体外构建工程化心肌组织的可行性。方法用含10μmol／L5-氮胞苷（5-Aza）的完全培养液（LG-DMEM，10％胎牛血清，100U／mL青霉素，100μg／mL链霉素，10μg／L碱性成纤维细胞生长因子）孵育第3代BMSCs24h后改用完全培养基培养4周，采用未处理组作为阴性对照。采用免疫细胞化学方法检测心肌肌钙蛋白T（cTnT）、α-肌动蛋白（α-Actin）的表达；逆转录-聚合酶链反应（RT-PCR）检测早期心肌形成转录因子GATA-4、Nkx2．5、TDGF-1基因的表达。将已诱导4周的BMSCs接种于多聚赖氨酸包被的脱细胞牛心包生物支架上培养2周后检测。结果诱导组细胞向心肌细胞转化，表达cTnT、α-Actin，对照组不表达cTnT、α-Actin；RT-PCR检测BMSCs诱导后表达早期心肌形成转录因子GATA-4、Nkx2．5、TDGF-1等基因；诱导后的BMSCs黏附于脱细胞牛心包生物支架表面，生长良好。结论大鼠BMSCs可向心肌细胞定向诱导分化，与脱细胞牛心包生物支架黏附良好，有望构建理想的组织工程化心肌。Objective To investigate the feasibility of inducing rat bone marrow mesenchymal stem cells （BMSCs）into myocardial ceils and the approach of applying the induced MSCs to construct myocardial tissue in vitro. Methods The third generation of BMSCs were incubated in complete culture medium （LGDMEM, 10%FBS, 100 U/mL penicillin, 100μg/mL streptomycin, 10 μg/L bFGF）, in which 10 μmol/L 5-Aza was added, for 24 hours. Then the BMSCs were switched to complete medium and cultured for 4 weeks as the experimental group. The other group without incubation was cultured in complete medium as the control, cTnT and a-Aetin were tested by immunocytochemistry method and the early myocardial transcription factor genes GATA- 4,Nkx2.5 ,TDGF-1 were tested by reverse transcription polymerase chain reaction（PT-PCR）. The induced BMSCs were planted onto the acellular bovine pericardium bio-stent which were coated by polylysine （PLYS） and cultured for 2 weeks before being tested. Results The experimental group showed morphological change to cardiomyocytes and expressed cTnT and a-Actin, while the control group did not express cTnT and α-Actin. The experimental group expressed the early myocardial transcription factor GATA-4, Nkx2.5, TDGF-1 gene. The induced BMSCs adhered to acellular bovine pericardium bio-stent surface and grew well. Conclusion BMSCs can be induced into cardiomyocyte in vitro and they adhere well with acellular bovine pericardium bio-stent. BMSCs have the potential to form an ideal cardiac muscle tissue.

关 键 词：骨髓间充质干细胞 心肌细胞 组织工程 

分 类 号：R54[医药卫生—心血管疾病]