Krack G, Gravier O, Roberfroid M, Mercier M
Biochim Biophys Acta. 1980 Nov 3;632(4):619-29. doi: 10.1016/0304-4165(80)90338-4.
An improved method for the homogenization and the subsequent subcellular fractionation of hepatocytes isolated from adult rat liver is described. The homogenization procedure developed in the present study allows the preservation of the integrity of subcellular structures, as demonstrated by measurement of the activities of representative enzymes as well as by determination of their latency. The activities of representative marker enzymes, as calculated on subcellular fractions obtained by differential centrifugation of the homogenate, are identical whether the homogenate arises from isolated hepatocytes or from the whole liver. Moreover, there is a close similitude between the kinetic parameters (Km and V) of two microsomal cytochrome P450-dependent mixed-function oxidases, namely aniline hydroxylase and aminopyrine demethylase determined on microsomal preparations obtained either from isolated cells or from the whole liver.
本文描述了一种改进的方法，用于对从成年大鼠肝脏中分离出的肝细胞进行匀浆及随后的亚细胞分级分离。本研究中开发的匀浆程序能够保持亚细胞结构的完整性，这一点通过测定代表性酶的活性以及确定其潜伏性得以证明。通过对匀浆进行差速离心获得的亚细胞级分计算得出的代表性标记酶活性，无论匀浆来自分离的肝细胞还是整个肝脏，都是相同的。此外，在分别从分离的细胞或整个肝脏获得的微粒体制剂上测定的两种微粒体细胞色素P450依赖性混合功能氧化酶，即苯胺羟化酶和氨基比林脱甲基酶的动力学参数（Km和V）之间存在密切相似性。
