摘要
目的探索较大尺度工程化肝组织原位肝表面贴覆片状植入、尽快建立血供的可行性。方法将采用胰酶冷消化法分离的新生大鼠肝细胞（1×10^10个／L）、肺组织块贴壁法分离的大鼠微血管内皮细胞（1×10^10个／L），与片状胶原支架、血管内皮生长因子（VEGF，10g／L）复合构建肝细胞／内皮细胞／血管生长因子／胶原支架复合的片状工程化肝组织。将其在15只大鼠肝表面贴覆植入，4周后取样进行大体观察及组织切片苏木素-伊红（HE）染色。结果分离的肝细胞及内皮细胞活率均为90％以上。片状工程化肝组织经肝表面贴覆法植入后，大体观察可见能与原位肝脏紧密贴合生长，形成体积大于1cm3的类肝样组织。体内植入后4周，植入物与肝脏贴合紧密，融合生长。组织学观察显示移植物中肝细胞均匀分布，未见细胞变性及死亡，有血管生成。结论肝表面贴覆植入法简便安全，可构建较大尺寸的工程化肝组织，有望达到原位修复受损肝脏的目的。
Objective To explore the feasibility of orthotopically implanting larger-size, sheet-like engineered liver tissue on the surface of liver to facilitate the establishment of blood supply. Methods Hepatocytes and microvessel endothelial cells isolated from newborn rats using enzymatic dissociation with typsin and by tissue adherence method, respectively, were combined with sheet-like collagen scaffold and vascular endothelial growth factor （VEGF, 10 g/L） to generate hepatocytes/endotheliocytes/VEGF/colla- gen constructs. The constructs were then implanted onto the surface of 15 rats＇ livers through the adherence method of sheet-like engineered liver tissue. Gross and histological observation was taken four weeks after implantation. Results The viability of both hepatocytes and endotheliocytes was over 90%. Sheet engi- neering liver tissue can fit closely with the orthotopic liver by the liver surface pasting method, and form hepatocyte-like tissue which was larger than 1 cm3. Hematoxylin and eosin （HE） staining showed that the hepatocytes were in good condition and distributed evenly in the implants, and angiogenesis could also he seen. We didn＇ t find cell degeneration and death. Conclusion The liver surface pasting method is sim- ple and safe, which can he used to build larger size of the engineering liver tissue, and also expected to reach the purpose of repairing damaged liver in situ.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第5期880-882,F0003,共4页
Chinese Journal of Experimental Surgery
基金
国家高技术研究发展计划（863计划）资助项目（2009A-A043801）
关键词
肝组织工程
新生大鼠肝细胞
血管内皮细胞
胶原支架材料
Liver tissue engineering
Newborn rat hepatocytes
Vascular endothelial cells
Collagen tissue scaffold
