A high-throughput cell-based toxicity analysis of drug metabolites using flow cytometry.
The effects of liver enzymes on drug activities are important considerations in the drug discovery process. Frequently, liver microsomes are used to simulate first-pass metabolism in the liver; however, there are significant disadvantages to the microsome system. As an alternative, a simple cell-based, high-throughput system that allows for examination of metabolite activity is described. Using multiparameter flow cytometry and the low-volume, high-sample format of 96-well plates, it is possible to rapidly evaluate a dose-response curve for metabolites based on variables including initial compound concentrations, hepatocyte cell line metabolic activities, and time. Using HepG2 cells as a surrogate for hepatic metabolism of a potential therapeutic, the impact of metabolites on Jurkat cell death was measured by both propidium iodide dye exclusion and cell cycle analysis. While this system is not proposed to supplant liver microsome studies, this alternative assay provides a highly adaptable, low-cost, and high-throughput measure of drug metabolism.
Buenz
E J
EJ
BioSciential LLC, Rochester, Minnesota 55903, USA. buenz@biosciential.com
D002232:Carbocyanines / Q000737:chemistry; D002453:Cell Cycle / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005434:Flow Cytometry / Q000379:methods*; D022781:Hepatocytes / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D019684:Magnoliopsida / Q000737:chemistry*; D010936:Plant Extracts / Q000378:metabolism*
D002232:Carbocyanines; D010936:Plant Extracts; C098044:3,3'-dioctadecyloxacarbocyanine