Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease.
Isolation, culture, and functional analysis of hepatocytes from mice with fatty liver disease.
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DOI：
10.1016/j.xpro.2020.100222
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发表时间：
2020-12-18
期刊：
影响因子：
--
通讯作者：
Svensson KJ
中科院分区：
文献类型：
--
作者：
Jung Y;Zhao M;Svensson KJ
我们提出了一个协议，用于分离肝细胞与建立非酒精性脂肪性肝病的小鼠。该方案包括使用蠕动泵进行肝脏灌注，然后采用改良的25%和90% Percoll梯度离心方案，以捕获通常使用传统分离方案丢失的载脂肝细胞。该方案能够同时分离正常和充满脂质的肝细胞。充满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。有关本方案使用和执行的完整详细信息，请参见和。该方案描述了从患有脂肪肝疾病的小鼠中分离肝细胞Percoll梯度离心能够进行基于密度的肝细胞分离提供了肝细胞脂质含量异质性的示例我们提出了一种从患有非酒精性脂肪肝疾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，然后采用改良的25%和90% Percoll梯度离心方案，以捕获通常使用传统分离方案丢失的载脂肝细胞。该方案能够同时分离正常和充满脂质的肝细胞。充满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。
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 and. This protocol describes the isolation of hepatocytes from mice with fatty liver disease Percoll gradient centrifugation enables density-based hepatocyte isolation Examples of heterogeneity in hepatocyte lipid content are presented 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.
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DOI：
10.1007/978-1-59745-019-5_13
发表时间：
2010-01-01
期刊：
MOUSE CELL CULTURE: METHODS AND PROTOCOLS
影响因子：
--
作者：
Li, Wan-Chun;Ralphs, Kate L.;Tosh, David
通讯作者：
Tosh, David
影响因子：
1.2
作者：
Kegel, Victoria;Deharde, Daniela;Damm, Georg
通讯作者：
Damm, Georg
DOI：
10.1007/978-1-62703-125-7_17
发表时间：
2013
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Shulman, Maria;Nahmias, Yaakov
通讯作者：
Nahmias, Yaakov
影响因子：
4.8
作者：
Radhakrishnan, Sridhar;Ke, Jia-Yu;Pellizzon, Michael A.
通讯作者：
Pellizzon, Michael A.
影响因子：
64.5
作者：
Sharabi K;Lin H;Tavares CDJ;Dominy JE;Camporez JP;Perry RJ;Schilling R;Rines AK;Lee J;Hickey M;Bennion M;Palmer M;Nag PP;Bittker JA;Perez J;Jedrychowski MP;Ozcan U;Gygi SP;Kamenecka TM;Shulman GI;Schreiber SL;Griffin PR;Puigserver P
通讯作者：
Puigserver P