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Abstract

Early stage pharmacological studies rely on in vitromethodologies for screening and testingcompounds. Conventional assays based on endpoint measurements provide limited informationbecause the lack in temporal resolution may not determine the pharmacological efect at its maximum.We developed an on-line, automated system for near real-time monitoring of extracellular contentfrom fve parallel suspension cultures, combining cell density measurements with a high-resolutionseparations every 12 minutes for 4 days. Selector and switching valves provide the fluidic controlrequired to sample from one culture during the analysis of the previous sample from another culture, atime-saving measure that is fundamental to the throughput of the presented system. The system wasapplied to study the metabolic efects of the drugs rotenone, β-lapachone and clioquinol using lactateas metabolic indicator. For each drug, 96 assays were executed on the extracellular matrix at threeconcentrations with two controls in parallel, consuming only 5.78mL of media from each culture overfour days, less than 60μL per analysis. The automated system provides high sample throughput, goodtemporal resolution and low sample consumption combined with a rugged analytical method withadequate sensitivity, providing a promising new platform for pharmacological and biotechnologicalstudies.

Item Type:	Article
Authors/Creators:	Alhusban, A and Breadmore, MC and Gueven, N and Guijt, RM
Journal or Publication Title:	Scientific Reports
Publisher:	Nature Publishing Group
ISSN:	2045-2322
DOI / ID Number:	10.1038/s41598-017-10472-1
Copyright Information:	Copyright 2017 The Authors. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) https://creativecommons.org/licenses/by/4.0/
Related URLs:	Google Scholar: Breadmore, MC UTAS Profile: Breadmore, MC Google Scholar: Gueven, N UTAS Profile: Gueven, N Google Scholar: Guijt, RM UTAS Profile: Guijt, RM
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