Li Wan-Chun, Ralphs Kate L, Tosh David
Department of Biology and Biochemistry, Centre for Regenerative Medicine, University of Bath, BA2 7AY Bath, UK.
Methods Mol Biol. 2010;633:185-96. doi: 10.1007/978-1-59745-019-5_13.
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.
肝脏具有多种功能，包括调节碳水化合物、脂肪和蛋白质代谢，对内源性和外源性生物活性物质进行解毒，以及合成和分泌血浆蛋白与胆汁。分离的肝细胞是在体外研究肝功能的一种有用技术。在此，我们描述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，即先用乙二胺四乙酸然后用胶原酶对肝脏进行连续灌注。分离后，细胞既可以用于短期研究，也可以在培养中长时间维持，以研究基因表达的长期变化。小鼠肝细胞分离方案可应用于正常小鼠和转基因小鼠。
