Abstract

Raman spectroscopy is successfully becoming an analytical tool used to characterize alterations in the biochemical composition of cells. In this work, we identify the features of Raman spectra of murine primary endothelial cells (EC) isolated from lungs, heart, liver, brain, kidney and aorta of normal mice, as well as from heart, lung and liver in a murine model of heart failure (HF) in Tgαq*44 mice. Primary cells were measured in suspension immediately after their isolation. Raman images showed that isolated primary EC were elliptical or circular, and did not show organ-specific spectral features for any of the studied organ, i.e. lungs, heart, liver, brain, kidney and aorta. Principal Component Analysis pairwise analysis of primary endothelial cells from FVB mice and Tgαq*44 mice revealed an increased protein content in EC isolated from the heart and increased lipid content in EC isolated from the lung in Tgαq*44 mice. No significant differences were found in the EC isolated from the liver using the same chemometric procedure. To our knowledge, this is the first report in which Raman spectroscopy has been used to characterize the biochemical phenotype of primary murine EC with developing HF. This pilot study shows that Raman-based analysis of freshly isolated primary EC did not revealed organ-specific features, however disease-associated changes were found in the coronary and pulmonary EC in the early stage of heart failure in Tgαq*44 mice.

Highlights

  • Endothelial cells (EC) form a unique monolayer that covers the luminal surface of all blood vessels maintaining cardiovascular ho­ meostasis

  • EC were isolated from the heart, aorta, lung, liver, kidney, and brain of FVB mice

  • The percentage of isolated CD31+/CD144+ cells ranged from 45%, 55%, 65%, 65%, 75% heart, up to 85%

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Summary

Introduction

Endothelial cells (EC) form a unique monolayer that covers the luminal surface of all blood vessels maintaining cardiovascular ho­ meostasis. It is a highly dynamic structure that is involved in the regulation of various processes including nutrient transport, vascular tone, inflammation, and thrombosis [1,2,3] Endothelium has both endocrine and paracrine functions and is involved in response to various physical and chemical signals through biosynthesis of various bioactive compounds [2,4]. The aim of these experiments was to characterize their biochemical composition, to look for organ-specific as well as disease specific features detected by Raman imaging in the isolated primary murine EC measured in suspension immediately after their isolation For his proof-of-concept study we have chosen FVB and Tgαq*44 mice, the latter representing the unique murine model of chronic heart failure [19,20,21,22,23]

Animals
Cell isolation and sample preparation
Flow cytometry
Results and discussion
Conclusions
Full Text
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