三种大鼠骨髓间充质干细胞源性人工肝组织的构建与评价  

Three kinds of engineered hepatic tissues constructed using rat bone marrow mesenchymal stem cells

作　　者：俞建雄[1] 袁静[2] 许瑞云[3] 

机构地区：[1]武汉大学人民医院胃肠外科,湖北省武汉市430060 [2]武汉大学人民医院眼科中心,湖北省武汉市430060 [3]中山大学附属第三医院肝胆外科,广东省广州市510630

出　　处：《中国组织工程研究》2014年第41期6578-6584,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81100664);武汉市青年科技晨光计划项目(2014070404010222);武汉大学自主科研基金资助项目(2042014kf0259)~~

摘　　要：背景:人工肝组织有可能为治疗急性肝衰竭提供一种新的选择,但人工肝组织的构建存在着种子细胞、支架、培养模式及对人工肝组织营养获取调控等各方面的难题。目的:体外构建3种人工肝组织,筛选最佳人工肝组织以供移植用。方法:纯化、扩增并诱导分化骨髓间充质干细胞为肝样细胞,在Transwell培养体系中分别与去细胞羊膜、DACRON PATCH心血管外科补片、生物外科补片3种支架结合构建3种人工肝组织,培养3 d后进行形态学和功能学检测。结果与结论:密度梯度离心结合贴壁法可获得较高纯度的大鼠骨髓间充质干细胞,并在体外转分化为肝样细胞。在构建的3种人工肝组织中,肝样细胞与生物外科补片支架材料可获得最大程度结合,并具有更强的尿素合成能力和白蛋白分泌功能,为治疗急性肝衰竭提供了实验基础。BACKGROUND：Engineered hepatic tissue is considered a promising strategy for healing acute liver failure. But, there are series of hindrances in the construction of engineered hepatic tissues, including acquisition of vital hepatocytes, choice of scaffolds and culture system, and nutrition supply. OBJECTIVE：To construct three kinds of engineered hepatic tissues in hope to screen the optimal one for transplantation in acute liver failure. METHODS：After purification, amplification, bone marrow mesenchymal stem cells were induced to differentiate into hepatocyte-like cells which were co-cultured with acellular amniotic membrane, DACRON PATCH cardiovascular surgical patch, biological surgical patch, respectively to construct three kinds of engineered hepatic tissues. After 3 days of culture, morphological and functional detections were performed. RESULTS AND CONCLUSION：Rat bone marrow mesenchymal stem cells with higher purity were successful y harvested by using density gradient centrifugation and adherent methods, and then the cells were differentiated into hepatocyte-like cells. In the three kinds of engineered hepatic tissues, hepatocyte-like cells were found to be combined with the biological surgical patch to the maximum extent, and their combination exhibited stronger ability of urea synthesis and albumin secretion, which provides experimental basis for treatment of acute liver failure.

关 键 词：干细胞 骨髓干细胞 急性肝衰竭 骨髓间充质干细胞 人工肝组织 Transwel 系统 羊膜 DACRON PATCH心血管外科补片 生物外科补片 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]