Metabolic fingerprints of human primary endothelial and fibroblast cells.

Polona Žigon,Snezna Sodin-Semrl,Saša Čučnik,Katjuša Mrak-Poljšak,Katja Lakota,Matija Tomsic,Matic Terčelj

doi:10.1007/s11306-016-1024-7

Abstract

IntroductionHuman primary cells originating from different locations within the body could differ greatly in their metabolic phenotypes, influencing both how they act during physiological/pathological processes and how susceptible/resistant they are to a variety of disease risk factors. A novel way to monitor cellular metabolism is through cell energetics assays, so we explored this approach with human primary cell types, as models of sclerotic disorders.ObjectivesIn order to better understand pathophysiological processes at the cellular level, our goals were to measure metabolic pathway activities of endothelial cells and fibroblasts, and determine their metabolic phenotype profiles.MethodsBiolog Phenotype MicroArray™ technology was used for the first time to characterize metabolic phenotypes of diverse primary cells. These colorimetric assays enable detection of utilization of 367 specific biochemical substrates by human endothelial cells from the coronary artery (HCAEC), umbilical vein (HUVEC) and normal, healthy lung fibroblasts (NHLF).ResultsAdenosine, inosine, d-mannose and dextrin were strongly utilized by all three cell types, comparable to glucose. Substrates metabolized solely by HCAEC were mannan, pectin, gelatin and prevalently tricarballylic acid. HUVEC did not show any uniquely metabolized substrates whereas NHLF exhibited strong utilization of sugars and carboxylic acids along with amino acids and peptides.ConclusionTaken together, we show for the first time that this simple energetics assay platform enables metabolic characterization of primary cells and that each of the three human cell types examined gives a unique and distinguishable profile.Electronic supplementary materialThe online version of this article (doi:10.1007/s11306-016-1024-7) contains supplementary material, which is available to authorized users.

Highlights

Introduction Human primary cells originating from different locations within the body could differ greatly in their metabolic phenotypes, influencing both how they act during physiological/pathological processes and how susceptible/resistant they are to a variety of disease risk factors
A novel way to monitor cellular metabolism is through cell energetics assays, so we explored this approach with human primary cell types, as models of sclerotic disorders
Biolog Phenotype MicroArrayTM technology was used for the first time to characterize metabolic phenotypes of diverse primary cells

Summary

Introduction

Human primary cells originating from different locations within the body could differ greatly in their metabolic phenotypes, influencing both how they act during physiological/pathological processes and how susceptible/resistant they are to a variety of disease risk factors. Cardiovascular cells that contribute directly to atherosclerosis and cardiac dysfunction are known to exhibit metabolic flexibility, characterized by the ability to switch from generating ATP primarily through oxidative phosphorylation to using glycolysis as the predominant energy source, as well as to shift from one fuel source to another (Vallerie and Bornfeldt 2015) This flexibility occurs in endothelial cells, myeloid cells, and cardiomyocytes during normal development and physiology, and is thought to have evolved to protect cells with heightened energy demand from the increased oxidative stress that can be a result of elevated rates of oxidative phosphorylation (Galvan-Pena and O’Neill 2014). Studies support the concept that metabolic flexibility confers the advantage of ensuring ATP supplies for continual cardiac contraction under a variety of physiological conditions (Goodwin and Taegtmeyer 2000; Kaijser and Berglund 1992; Schonekess 1997; Wentz et al 2010)

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