Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells, and Kupffer Cells
Advanced Method for Isolation of Mouse Hepatocytes, Liver Sinusoidal Endothelial Cells, and Kupffer Cells
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DOI：
10.1007/978-1-4939-6700-1_21
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发表时间：
2017-01-01
期刊：
影响因子：
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通讯作者：
Lu, Mengji
中科院分区：
文献类型：
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作者：
Liu, Jia;Huang, Xuan;Lu, Mengji
从肝脏分离纯细胞群是研究肝实质细胞和非实质细胞在肝脏生理学、病理生理学和免疫学中的作用的先决条件。传统的肝细胞分离方法通常只从肝标本中纯化单一类型的细胞。在这里，我们描述了一种有效的方法，可以同时从单个小鼠肝脏标本中纯化肝细胞(HC)、肝窦内皮细胞(LSECs)和库普弗细胞(KCs)。以解放酶为基础的灌流技术结合低速离心和磁激活细胞分选(MACS)技术，高产率和高纯度地分离和纯化了HC、KCs和LSECs。
Separation of pure cell populations from the liver is a prerequisite to study the role of hepatic parenchymal and non-parenchymal cells in liver physiology, pathophysiology, and immunology. Traditional methods for hepatic cell separation usually purify only single cell types from liver specimens. Here, we describe an efficient method that can simultaneously purify populations of hepatocytes (HCs), liver sinusoidal endothelial cells (LSECs), and Kupffer cells (KCs) from a single mouse liver specimen. A liberase-based perfusion technique in combination with a low-speed centrifugation and magnetic-activated cell sorting (MACS) led to the isolation and purification of HCs, KCs, and LSECs with high yields and purity.