Isolation and Culture of Adult Mouse Hepatocytes
Isolation and Culture of Adult Mouse Hepatocytes
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DOI：
10.1007/978-1-59745-019-5_13
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Tosh, David
中科院分区：
文献类型：
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作者：
Li, Wan-Chun;Ralphs, Kate L.;Tosh, David
肝脏具有多种功能，包括调节碳水化合物、脂肪和蛋白质的代谢，体内和外源生物的解毒，以及血浆蛋白和胆汁的合成和分泌。分离的肝细胞是在体外研究肝功能的一种有用的技术。本文描述了一种从成年小鼠肝脏中分离肝细胞的方法。该方法的原理是两步胶原酶灌注技术，包括用乙二胺四乙酸和胶原酶依次灌注肝脏。分离后，细胞既可用于短期研究，也可在培养中长期保存以研究基因表达的长期变化。小鼠肝细胞分离方案可适用于正常小鼠和转基因小鼠。
The liver performs a multitude of functions including the regulation of carbohydrate, fat, and protein metabolism, the detoxification of endo- and xenobiotics, and the synthesis and secretion of plasma proteins and bile. Isolated hepatocytes constitute a useful technique for studying liver function in an in vitro setting. Here we describe a method for the isolation of hepatocytes from adult mouse liver. The principle of the method is the two-step collagenase perfusion technique which involves sequential perfusion of the liver with ethylenediaminetetraacetic acid and collagenase. Following isolation, the cells can either be used for short-term studies or, alternatively, maintained in culture for prolonged periods to study long-term changes in gene expression. The protocol for mouse hepatocyte isolation may be applied to both normal and transgenic mice.