Acute Metabolic Switch Assay Using Glucose/Galactose Medium in HepaRG Cells to Detect Mitochondrial Toxicity
Laleh Kamalian, Oisin Douglas, Carol E. Jolly, Jan Snoeys, Damir Simic, Mario Monshouwer, Dominic P. Williams, B. Kevin Park, Amy E. Chadwick
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引用次数: 9
Abstract
Using galactose instead of glucose in the culture medium of hepatoma cell lines, such as HepG2 cells, has been utilized for a decade to unmask the mitochondrial liability of chemical compounds. A modified glucose-galactose assay on HepG2 cells, reducing the experimental period for screening of mitochondrial toxicity to 2 to 4 hr, has been previously reported. HepaRG cells are one of the few cell lines that retain some of the important characteristics of human hepatocytes, offering advantages of working with a cell line, therefore, are considered an alternative for HepG2 cells in drug toxicity screening. A method is described here using HepaRG cells in an acute metabolic switch assay utilizing specific glucose/galactose media, a combined ATP-protein-LDH assay measuring three endpoints from one 96-well plate, and a criteria to label a compound as a mitochondrial toxin. © 2019 by John Wiley & Sons, Inc.
使用葡萄糖/半乳糖培养基检测HepaRG细胞线粒体毒性的急性代谢开关试验
在肝癌细胞系(如HepG2细胞)的培养基中使用半乳糖代替葡萄糖,已经使用了十年,以揭示化学化合物的线粒体倾向。先前有报道称,在HepG2细胞上进行了一种改进的葡萄糖-半乳糖试验,将线粒体毒性筛选的实验时间缩短至2至4小时。HepaRG细胞是少数保留人类肝细胞一些重要特征的细胞系之一,具有与细胞系一起工作的优势,因此被认为是HepG2细胞在药物毒性筛选中的替代品。本文描述了一种方法,使用HepaRG细胞进行急性代谢开关试验,利用特定的葡萄糖/半乳糖培养基,结合atp蛋白- ldh试验,测量一个96孔板的三个端点,以及将化合物标记为线粒体毒素的标准。©2019 by John Wiley &儿子,Inc。
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