Metabolomics guides rational development of a simplified cell culture medium for drug screening against Trypanosoma brucei

Creek, D. J., Nijagal, B., Kim, D.-H., Rojas, F., Matthews, K. R. and Barrett, M. P. (2013) Metabolomics guides rational development of a simplified cell culture medium for drug screening against Trypanosoma brucei. Antimicrobial Agents and Chemotherapy, 57(6), pp. 2768-2779. (doi:10.1128/AAC.00044-13)

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Abstract

n vitro culture methods underpin many experimental approaches to biology and drug discovery. The modification of established cell culture methods to make them more biologically relevant or to optimize growth is traditionally a laborious task. Emerging metabolomic technology enables the rapid evaluation of intra- and extracellular metabolites and can be applied to the rational development of cell culture media. In this study, untargeted semiquantitative and targeted quantitative metabolomic analyses of fresh and spent media revealed the major nutritional requirements for the growth of bloodstream form Trypanosoma brucei. The standard culture medium (HMI11) contained unnecessarily high concentrations of 32 nutrients that were subsequently removed to make the concentrations more closely resemble those normally found in blood. Our new medium, Creek's minimal medium (CMM), supports in vitro growth equivalent to that in HMI11 and causes no significant perturbation of metabolite levels for 94% of the detected metabolome (<3-fold change; α = 0.05). Importantly, improved sensitivity was observed for drug activity studies in whole-cell phenotypic screenings and in the metabolomic mode of action assays. Four-hundred-fold 50% inhibitory concentration decreases were observed for pentamidine and methotrexate, suggesting inhibition of activity by nutrients present in HMI11. CMM is suitable for routine cell culture and offers important advantages for metabolomic studies and drug activity screening.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Barrett, Professor Michael and Creek, Dr Darren and Rojas, Mr Federico and Nijagal, Ms Brunda
Authors: Creek, D. J., Nijagal, B., Kim, D.-H., Rojas, F., Matthews, K. R., and Barrett, M. P.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Antimicrobial Agents and Chemotherapy
Journal Abbr.:Antimicrob Agents Chemother
Publisher:American Society for Microbiology
ISSN:0066-4804
ISSN (Online):1098-6596
Copyright Holders:Copyright © 2013 American Society for Microbiology
First Published:First published in Antimicrobial Agents and Chemotherapy 57(6):2768-2779
Publisher Policy:Reproduced in accordance with the copyright policy of the publisher
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
371796The Wellcome Centre for Molecular Parasitology ( Core Support )J BarryWellcome Trust (WELLCOME)085349/Z/08/ZIII - BACTERIOLOGY