Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures

Abstract Early stage pharmacological studies rely on in vitro methodologies for screening and testing compounds. Conventional assays based on endpoint measurements provide limited information because the lack in temporal resolution may not determine the pharmacological effect at its maximum. We deve...

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Autores principales:	Ala A. Alhusban, Michael C. Breadmore, Nuri Gueven, Rosanne M. Guijt
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/aba12791db91466da529719d8df4d111
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spelling	oai:doaj.org-article:aba12791db91466da529719d8df4d1112021-12-02T15:04:56ZTime-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures10.1038/s41598-017-10472-12045-2322https://doaj.org/article/aba12791db91466da529719d8df4d1112017-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-10472-1https://doaj.org/toc/2045-2322Abstract Early stage pharmacological studies rely on in vitro methodologies for screening and testing compounds. Conventional assays based on endpoint measurements provide limited information because the lack in temporal resolution may not determine the pharmacological effect at its maximum. We developed an on-line, automated system for near real-time monitoring of extracellular content from five parallel suspension cultures, combining cell density measurements with a high-resolution separations every 12 minutes for 4 days. Selector and switching valves provide the fluidic control required to sample from one culture during the analysis of the previous sample from another culture, a time-saving measure that is fundamental to the throughput of the presented system. The system was applied to study the metabolic effects of the drugs rotenone, β-lapachone and clioquinol using lactate as metabolic indicator. For each drug, 96 assays were executed on the extracellular matrix at three concentrations with two controls in parallel, consuming only 5.78 mL of media from each culture over four days, less than 60 μL per analysis. The automated system provides high sample throughput, good temporal resolution and low sample consumption combined with a rugged analytical method with adequate sensitivity, providing a promising new platform for pharmacological and biotechnological studies.Ala A. AlhusbanMichael C. BreadmoreNuri GuevenRosanne M. GuijtNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ala A. Alhusban Michael C. Breadmore Nuri Gueven Rosanne M. Guijt Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures
description	Abstract Early stage pharmacological studies rely on in vitro methodologies for screening and testing compounds. Conventional assays based on endpoint measurements provide limited information because the lack in temporal resolution may not determine the pharmacological effect at its maximum. We developed an on-line, automated system for near real-time monitoring of extracellular content from five parallel suspension cultures, combining cell density measurements with a high-resolution separations every 12 minutes for 4 days. Selector and switching valves provide the fluidic control required to sample from one culture during the analysis of the previous sample from another culture, a time-saving measure that is fundamental to the throughput of the presented system. The system was applied to study the metabolic effects of the drugs rotenone, β-lapachone and clioquinol using lactate as metabolic indicator. For each drug, 96 assays were executed on the extracellular matrix at three concentrations with two controls in parallel, consuming only 5.78 mL of media from each culture over four days, less than 60 μL per analysis. The automated system provides high sample throughput, good temporal resolution and low sample consumption combined with a rugged analytical method with adequate sensitivity, providing a promising new platform for pharmacological and biotechnological studies.
format	article
author	Ala A. Alhusban Michael C. Breadmore Nuri Gueven Rosanne M. Guijt
author_facet	Ala A. Alhusban Michael C. Breadmore Nuri Gueven Rosanne M. Guijt
author_sort	Ala A. Alhusban
title	Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures
title_short	Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures
title_full	Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures
title_fullStr	Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures
title_full_unstemmed	Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures
title_sort	time-resolved pharmacological studies using automated, on-line monitoring of five parallel suspension cultures
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/aba12791db91466da529719d8df4d111
work_keys_str_mv	AT alaaalhusban timeresolvedpharmacologicalstudiesusingautomatedonlinemonitoringoffiveparallelsuspensioncultures AT michaelcbreadmore timeresolvedpharmacologicalstudiesusingautomatedonlinemonitoringoffiveparallelsuspensioncultures AT nurigueven timeresolvedpharmacologicalstudiesusingautomatedonlinemonitoringoffiveparallelsuspensioncultures AT rosannemguijt timeresolvedpharmacologicalstudiesusingautomatedonlinemonitoringoffiveparallelsuspensioncultures
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Time-Resolved Pharmacological Studies using Automated, On-line Monitoring of Five Parallel Suspension Cultures

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