Long-term culture and coculture of primary rat and human hepatocytes.
Long-term culture and coculture of primary rat and human hepatocytes.
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DOI：
10.1007/978-1-62703-125-7_17
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Nahmias, Yaakov
中科院分区：
文献类型：
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作者：
Shulman, Maria;Nahmias, Yaakov
肝脏是哺乳动物最大的内脏器官，具有广泛的生命功能。在美国，由于药物毒性或病毒感染导致的肝功能丧失是死亡的主要原因。生物人工肝（BAL）装置的发展以及药物和化妆品毒性筛选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迅速丧失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的现象。近年来，开发了几种技术，可以在培养物中支持高水平的肝脏特异性基因表达、代谢和合成功能。这些包括胶原双凝胶构型、肝细胞球状体、与内皮细胞共培养以及与3 T3-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-细胞相互作用，和基本功能测定。
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.