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Bioanalysis. 2013 Dec;5(24):3009-21. doi: 10.4155/bio.13.269.

Human platelets as a platform to monitor metabolic biomarkers using stable isotopes and LC-MS.

Author information

1
Department of Pharmacology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.

Abstract

BACKGROUND:

Intracellular metabolites such as CoA thioesters are modulated in a number of clinical settings. Their accurate measurement from surrogate tissues such as platelets may provide additional information to current serum and urinary biomarkers.

METHODS:

Freshly isolated platelets from healthy volunteers were treated with rotenone, propionate or isotopically labeled metabolic tracers. Using a recently developed LC-MS-based methodology, absolute changes in short-chain acyl-CoA thioesters were monitored, as well as relative metabolic labeling using isotopomer distribution analysis.

RESULTS:

Consistent with in vitro experiments, isolated platelets treated with rotenone showed decreased intracellular succinyl-CoA and increased β-hydroxybutyryl-CoA, while propionate treatment resulted in increased propionyl-CoA. In addition, isotopomers of the CoAs were readily detected in platelets treated with the [(13)C]- or [(13)C(15)N]-labeled metabolic precursors.

CONCLUSION:

Here, we show that human platelets can provide a powerful ex vivo challenge platform with potential clinical diagnostic and biomarker discovery applications.

PMID:
24320127
PMCID:
PMC3912754
DOI:
10.4155/bio.13.269
[Indexed for MEDLINE]
Free PMC Article
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