Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA.
Stanford Diabetes Research Center, Stanford University School of Medicine, Stanford, CA 94305, USA.
STAR Protoc. 2020 Dec 15;1(3):100222. doi: 10.1016/j.xpro.2020.100222. eCollection 2020 Dec 18.
We present a protocol for isolating hepatocytes from mice with established non-alcoholic fatty liver disease. This protocol consists of liver perfusion using a peristaltic pump, followed by a modified 25% and 90% Percoll gradient centrifugation protocol to capture lipid-laden hepatocytes that are usually lost using traditional isolation protocols. This protocol enables simultaneous isolation of normal and lipid-filled hepatocytes. Lipid-filled hepatocytes can be used in cell culture systems to study drug metabolism, hepatotoxicity, or glucose and lipid metabolism. For complete details on the use and execution of this protocol, please refer to Sharabi et al. (2017) and Kegel et al. (2016).
我们介绍了一种从患有非酒精性脂肪性肝病的小鼠中分离肝细胞的方案。该方案包括使用蠕动泵进行肝脏灌注，然后使用改良的 25%和 90%Percoll 梯度离心方案来捕获通常使用传统分离方案丢失的富含脂质的肝细胞。该方案能够同时分离正常和富含脂质的肝细胞。富含脂质的肝细胞可用于细胞培养系统中，以研究药物代谢、肝毒性或葡萄糖和脂质代谢。有关此方案的使用和执行的完整详细信息，请参阅 Sharabi 等人。（2017 年）和 Kegel 等人。（2016 年）。
