聚乳酸-聚乙醇酸共聚物复合心肌样细胞体外构建工程化心肌组织  

Construction of engineered myocardial tissues with polylactic-co-glycolic acid polymer and cardiomyocyte-like cells in vitro

作　　者：邢玉洁[1] 吕安林[1] 王利[1] 燕学波[1] 

机构地区：[1]解放军第四军医大学西京医院心血管内科,陕西省西安市710032

出　　处：《中国组织工程研究与临床康复》2010年第16期2875-2878,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:研究证实,在一定的诱导条件下,骨髓间充质干细胞可能向心肌细胞等多种中胚层来源的间质细胞分化。目的:探索以聚乳酸-聚乙醇酸共聚物为支架、骨髓间充质干细胞诱导分化的心肌样细胞为种子细胞,体外构建工程化心肌组织的可行性。方法:分离SD大鼠骨髓间充质干细胞,取第3代骨髓间充质干细胞加入含5-氮胞苷的培养液进行诱导分化,培养4周。将诱导成功后的细胞制成细胞悬液,缓慢滴注于预先制备好的聚乳酸-聚乙醇酸共聚物上,培养14d。以倒置相差显微镜观察诱导前后细胞的形态学变化,免疫荧光染色法鉴定诱导后骨髓间充质干细胞中心肌特异性肌钙蛋白Ⅰ的表达,大体观察工程化心肌组织在培养期间的形态,透射电镜观察工程化心肌组织的超微结构。结果与结论:原代培养的骨髓间充质干细胞14d形成集落,传代细胞体积变大,5-氮胞苷诱导后细胞呈长梭形,呈一致性生长。免疫荧光染色结果显示诱导后骨髓间充质干细胞表达心肌特异性肌钙蛋白Ⅰ。透射电镜可见肌丝、Z线样物质。结果证实,以聚乳酸-聚乙醇酸共聚物为支架、骨髓间充质干细胞诱导分化的心肌样细胞为种子细胞,可于体外构建出类似天然心肌的工程化心肌组织。BACKGROUND：Previous research has demonstrated that bone marrow mesenchymal stem cells（BMSCs）differentiate into varying mesoderm-derived mesenchymal cells,such as myocardial cells.OBJECTIVE：To investigate the feasibility of construction of engineered myocardial tissues with polylactic-co-glycolic acid（PLGA） polymer and cardiomyocyte-like cells derived from BMSCs in vitro.METHODS：BMSCs were isolated from bone marrow of SD rats by density gradient centrifugation.The third passage cells were induced with the culture medium including 5-aza for 4 weeks.After successful induction,the cells were trypsinized and suspended,then the cell suspension was added to the polylactic-co-glycolic acid square slowly and cultured in the incubator for 14 days.The morphological changes were observed before and after induction under phase contrast microscope.The cardiomyocyte-like cells were identified by immunofluorescence staining.The naked eyes were used to observe the morphology of engineered myocardial tissues.The ultrastructures of the engineered myocardial tissues were viewed with a transmission electron microscope.RESULTS AND CONCLUSION：BMSCs of primary culture formed cell colonies at 14 days.The passaged cells were larger than those of primary culture.After induction by 5-aza,the cells presented long spindle and aligned in parallel.The expression of specific proteins of cardiac troponin I（cTnI）in induced BMSCs was positive.Transmission electron microscopy showed that the engineered myocardial tissues had myofilaments,Z line-like substances.With PLGA and cardiomyocyte-like cells derived from BMSCs,the engineered myocardial tissues which resemble native cardiac tissues can be successfully constructed in vitro.

关 键 词：聚乳酸-聚乙醇酸 5-氮杂胞苷 骨髓间充质干细胞 心肌样细胞 工程化心肌组织 

分 类 号：R318[医药卫生—生物医学工程]