Abstract 407: Use of Micellarized Cholesteryl Ester to Generate Macrophage Foam Cells
Background Development of foam cells for Reverse Cholesterol Transport (RCT) studies uses the technique of receptor dependent macrophage loading with radiolabeled acetylated LDL. For various reasons this technique is tedious, less reliable and less reproducible. Our goal is to develop foam cells loaded Cholesteryl Ester (CE) in a receptor independent manner and facile manner.
Methods We micellarized highly nonpolar Cholesterol or CE in aqueous solution of Lyso phosphatidylcholine (Lyso PtdCho) to generate mixed micelles. RAW 264.7 macrophages and mouse peritoneal macrophages were incubated with Cholesterol or CE (unlabeled and fluorescently labeled-NBD cholesterol)/ Lyso PtdCho mixed micelles. The presence of fluorescent Cholesterol and CE was documented by fluorescence microscopy. Foam cell macrophages were detected by Oil-red O staining.
Results In the presence of Lyso PtdCho, Cholesterol or CE readily formed mixed micelles, generating a clear solution in a concentration-dependent manner. Incubation of Cholesterol or CE rich micelles with macrophages resulted in the uptake of these micelles as noted by fluorescence and Oil-red O staining.
Conclusions Foam cells developed by this technique could be used for RCT studies. This technique can be manipulated to enrich cells in vitro with various non-polar molecules and could be used to develop different cell based assays.
- © 2013 by American Heart Association, Inc.