Institute on Membrane Technology, National Research Council of Italy, ITM-CNR, c/o University of Calabria, Rende (CS), Italy.
Biomaterials. 2011 Dec;32(34):8848-59. doi: 10.1016/j.biomaterials.2011.08.004. Epub 2011 Aug 25.
The realization of organotypic liver model that exhibits stable phenotype is a major challenge in the field of liver tissue engineering. In this study we developed liver organotypic co-culture systems by using synthetic and biodegradable membranes with primary human hepatocytes and human umbilical vein endothelial cells (HUVEC). Synthetic membranes prepared by a polymeric blend constituted of modified polyetheretherketone (PEEK-WC) and polyurethane (PU) and biodegradable chitosan membranes were developed by phase inversion technique and used in homotypic and organotypic culture systems. The morphological and functional characteristics of cells in the organotypic co-culture membrane systems were evaluated in comparison with homotypic cultures and traditional systems. Hepatocytes in the organotypic co-culture systems exhibit compact polyhedral cells with round nuclei and well demarcated cell-cell borders like in vivo, as a result of heterotypic interaction with HUVECs. In addition HUVECs formed tube-like structures directly through the interactions with the membranes and hepatocytes and indirectly through the secretion of ECM proteins which secretion improved in the organotypic co-culture membrane systems. The heterotypic cell-cell contacts have beneficial effect on the hepatocyte albumin production, urea synthesis and drug biotransformation. The developed organotypic co-culture membrane systems elicit liver specific functions in vitro and could be applied for the realization of engineered liver tissues to be used in tissue engineering, drug metabolism studies and bioartificial liver devices.
实现具有稳定表型的器官型肝模型是肝组织工程领域的主要挑战。在这项研究中，我们使用原代人肝细胞和人脐静脉内皮细胞（HUVEC）开发了肝器官型共培养系统，使用了由改性聚醚醚酮（PEEK-WC）和聚氨酯（PU）组成的聚合物共混物以及通过相转化技术制备的可生物降解壳聚糖膜制备的合成膜，并用于同型和器官型培养系统。与同型培养和传统系统相比，评估了器官型共培养膜系统中细胞的形态和功能特征。器官型共培养系统中的肝细胞呈紧密的多面体细胞，细胞核呈圆形，细胞-细胞边界清晰，这是与 HUVEC 异型相互作用的结果。此外，HUVEC 直接通过与膜和肝细胞的相互作用形成管状结构，间接通过 ECM 蛋白的分泌来促进，而这种分泌在器官型共培养膜系统中得到了改善。异型细胞-细胞接触对肝细胞白蛋白产生、尿素合成和药物生物转化具有有益的影响。开发的器官型共培养膜系统在体外引起了肝脏的特异性功能，可用于实现用于组织工程、药物代谢研究和生物人工肝脏装置的工程化肝脏组织。
