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比较基因表达的 RAP-PCR 分析在新鲜和永生化大鼠肝细胞系。

Comparison of RAP-PCR analysis of gene expression in fresh and immortalised rat hepatocyte cell lines.

机构信息

Biological Sciences, University of Paisley, Paisley, UK.

出版信息

Cytotechnology. 2000 Oct;34(1-2):159-63. doi: 10.1023/A:1008159912634.

Abstract

Primary rat hepatocytes dedifferentiate rapidly losing theactivities of the drug metabolising enzymes involved in thedetoxification of xenobiotics in the liver. An alternativeapproach to using primary hepatocytes for toxicity testing isthe development of immortalised hepatocyte cell lines via thetransfection of primary hepatocytes with SV40 DNA. In order toassess the suitability of immortalised lines as an alternativeto primary cell cultures we have used RNA arbitrarily primedpolymerase chain reaction to compare gene expression inimmortalised rat hepatocyte cell lines with that in primary rathepatocytes. We have found that differences exist in the RNAtranscripts between fresh and immortalised hepatocyteshighlighting RNA arbitrarily primed polymerase chain reaction asa useful screening method for identifying immortalised lineswhich retain the most ;normal' phenotype in relation to theprimary cells from which they originated.

摘要

原代大鼠肝细胞迅速去分化,失去肝脏中参与解毒的药物代谢酶的活性。一种替代使用原代肝细胞进行毒性测试的方法是通过用 SV40 DNA 转染原代肝细胞来开发永生化的肝细胞系。为了评估永生化细胞系作为原代细胞培养物的替代物的适用性,我们使用 RNA 任意引物聚合酶链反应来比较永生化大鼠肝细胞系与原代大鼠肝细胞中的基因表达。我们发现新鲜和永生化肝细胞之间的 RNA 转录本存在差异,这突出显示了 RNA 任意引物聚合酶链反应是一种有用的筛选方法,可用于鉴定保留与原始细胞最相关的“正常”表型的永生化细胞系。

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