Shulman Maria, Nahmias Yaakov
The Selim and Rachel Benin School of Engineering, The Hebrew University of Jerusalem, Jerusalem, Israel.
Methods Mol Biol. 2013;945:287-302. doi: 10.1007/978-1-62703-125-7_17.
The liver is the largest internal organ in mammals, serving a wide spectrum of vital functions. Loss of liver function due to drug toxicity or viral infection is a major cause of death in the United States. The development of Bioartificial Liver (BAL) devices and the demand for pharmaceutical and cosmetic toxicity screening require the development of long-term hepatocyte culture techniques. However, primary hepatocytes rapidly lose their cuboidal morphology and liver-specific functions over a few days in culture. Accumulation of stress fibers, loss of metabolic function, and cell death are known phenomena. In recent years, several techniques were developed that can support high levels of liver-specific gene expression, metabolic and synthetic function for several weeks in culture. These include the collagen double-gel configuration, hepatocyte spheroids, coculture with endothelial cells, and micropatterned cocultures with 3T3-J2 fibroblasts. This chapter covers the current status of hepatocyte culture techniques, including: hepatocyte isolation, media formulation, oxygen supply, heterotypic cell-cell interactions, and basic functional assays.
肝脏是哺乳动物体内最大的内脏器官，具有多种重要功能。在美国，药物毒性或病毒感染导致的肝功能丧失是主要死因之一。生物人工肝（BAL）装置的研发以及药物和化妆品毒性筛查的需求，要求开发长期肝细胞培养技术。然而，原代肝细胞在培养几天后会迅速失去其立方形形态和肝脏特异性功能。应激纤维的积累、代谢功能的丧失和细胞死亡是已知现象。近年来，人们开发了几种技术，能够在培养数周内维持高水平的肝脏特异性基因表达、代谢和合成功能。这些技术包括胶原双凝胶结构、肝细胞球体、与内皮细胞共培养以及与3T3-J2成纤维细胞的微图案化共培养。本章涵盖了肝细胞培养技术的现状，包括：肝细胞分离、培养基配方、氧气供应、异型细胞间相互作用以及基本功能测定。
