Co-culture models of endothelial cells, macrophages, and vascular smooth muscle cells for the study of the natural history of atherosclerosis

• Published in: PloS one Vol. 18; no. 1; p. e0280385
• Main Authors: Liu, Martin, Samant, Saurabhi, Vasa, Charu Hasini, Pedrigi, Ryan M, Oguz, Usama M, Ryu, Sangjin, Wei, Timothy, Anderson, Daniel R, Agrawal, Devendra K, Chatzizisis, Yiannis S
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 20.01.2023; Public Library of Science (PLoS)
• Subjects: Arteriosclerosis; Atherogenesis; Atherosclerosis; Atherosclerosis - metabolism; Biology; Biology and Life Sciences; Care and treatment; Cell culture; Coculture Techniques; Collagen; Collagen (type I); Confocal microscopy; Coronary artery; Coronary vessels; Coupling (molecular); Diagnosis; Electron microscopy; Endothelial cells; Endothelial Cells - metabolism; Endothelium; Health aspects; Humans; Immunofluorescence; Inflammation; Lipids; Lipoproteins; Low density lipoprotein; Macrophages; Macrophages - metabolism; Medicine and Health Sciences; Microscopy; Modelling; Monocytes; mRNA; Muscle, Smooth, Vascular - metabolism; Muscles; Myocytes, Smooth Muscle - metabolism; Pathophysiology; Physical Sciences; Research and Analysis Methods; Shear stress; Smooth muscle; Tendons; Thickening; Veins & arteries; Xanthoma; United States; United States > US
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0280385

This work aims to present a fast, affordable, and reproducible three-cell co-culture system that could represent the different cellular mechanisms of atherosclerosis, extending from atherogenesis to pathological intimal thickening. We built four culture models: (i) Culture model #1 (representing normal arterial intima), where human coronary artery endothelial cells were added on top of Matrigel-coated collagen type I matrix, (ii) Culture model #2 (representing atherogenesis), which demonstrated the subendothelial accumulation and oxidative modification of low-density lipoproteins (LDL), (iii) Culture model #3 (representing intimal xanthomas), which demonstrated the monocyte adhesion to the endothelial cell monolayer, transmigration into the subendothelial space, and transformation to lipid-laden macrophages, (iv) Culture model #4 (representing pathological intimal thickening), which incorporated multiple layers of human coronary artery smooth muscle cells within the matrix. Coupling this model with different shear stress conditions revealed the effect of low shear stress on the oxidative modification of LDL and the upregulation of pro-inflammatory molecules and matrix-degrading enzymes. Using electron microscopy, immunofluorescence confocal microscopy, protein and mRNA quantification assays, we showed that the behaviors exhibited by the endothelial cells, macrophages and vascular smooth muscle cells in these models were very similar to those exhibited by these cell types in nascent and intermediate atherosclerotic plaques in humans. The preparation time of the cultures was 24 hours. We present three-cell co-culture models of human atherosclerosis. These models have the potential to allow cost- and time-effective investigations of the mechanobiology of atherosclerosis and new anti-atherosclerotic drug therapies.