Isolation of human fetal liver progenitors and their enhanced proliferation by three-dimensional coculture with endothelial cells.
Isolation of human fetal liver progenitors and their enhanced proliferation by three-dimensional coculture with endothelial cells.
复制标题
人胎肝祖细胞的分离及其通过与内皮细胞三维共培养增强增殖。
DOI：
10.1089/ten.tea.2007.0087
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发表时间：
2008
影响因子：
--
通讯作者：
Millan,MariaT
中科院分区：
文献类型：
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作者：
Xiong,Anming;Austin,TimothyW;Lagasse,Eric;Uchida,Nobuko;Tamaki,Stanley;Bordier,BrunoB;Weissman,IrvingL;Glenn,JeffreyS;Millan,MariaT
通过流式细胞术从原代人胎肝中分离肝祖细胞，其特征在于共表达胆汁和肝细胞谱系标记物以及在培养物中形成集落的能力。这些前瞻性分离的肝祖细胞支持丁型肝炎病毒感染，表达并产生白蛋白和甲胎蛋白，分别通过白蛋白和甲胎蛋白驱动的慢病毒启动子报告基因构建体进行跟踪，并通过ELISA进行测量。在三维（3D）纤维蛋白凝胶与内皮细胞的共培养导致血管结构的内皮细胞和肝祖细胞的增殖增加的形成。当肝祖细胞和内皮细胞直接接触培养时观察到的肝祖细胞增殖的增强，当肝祖细胞和内皮细胞在相邻但分开的基质上培养时以及当它们跨transwell膜培养时没有实现。总之，肝祖细胞和内皮细胞在三维基质中的共培养导致肝祖细胞增殖和功能增强。这种共培养方法提供了一种新的共培养系统，可以应用于工程肝组织的发展。
Liver progenitor cells, characterized by the coexpression of biliary and hepatocyte lineage markers and the ability to form colonies in culture, were isolated by flow cytometry from primary human fetal livers. These prospectively isolated liver progenitor cells supported hepatitis D virus infection, expressed, and produced albumin and α-fetoprotein, as tracked by albumin- and α-fetoprotein–driven lentiviral promoter reporter constructs and measured by ELISA, respectively. Coculture in three-dimensional (3D) fibrin gel with endothelial cells resulted in the formation of vascular structures by the endothelial cells and increased proliferation of liver progenitors. The enhanced proliferation of liver progenitors that was observed when liver progenitors and endothelial cells were cultured in direct contact was not achieved when liver progenitors and endothelial cells were cultured on adjacent but separate matrices and when they were cultured across transwell membranes. In conclusion, coculture of liver progenitors and endothelial cells in three-dimensional matrix resulted in enhanced liver progenitor proliferation and function. This coculture methodology offers a novel coculture system that could be applied for the development of engineered liver tissues.