Sampath Parthasarathy
University of Central Florida, USA
Title: Novel ways to lipid load cells to study reverse cholesterol transport.
Biography
Biography: Sampath Parthasarathy
Abstract
Generation of foam cells, an essential step for reverse cholesterol transport (RCT) studies, uses the technique of receptor dependent macrophage loading with radiolabeled acetylated Low Density Lipoprotein (Ac-LDL). In this study, we used the ability of a biologically relevant detergent molecule, Lysophosphatidylcholine (Lyso PtdCho), to form mixed micelles with cholesterol or cholesteryl ester (CE) to generate macrophage foam cells. Fluorescent or radiolabelled cholesterol / Lyso PtdCho mixed micelles were prepared and incubated with RAW 264.7 or mouse peritoneal macrophages. Macrophages incubated with cholesterol or CE (unlabeled, fluorescently labeled or radiolabeled) / Lyso PtdCho mixed micelles accumulated CE as documented by microscopy, lipid staining, labeled oleate incorporation, and by thin layer chromatography (TLC). Such foam cells unloaded cholesterol when incubated with high density lipoprotein (HDL) and not with oxidized HDL (Ox-HDL). We propose that stable cholesterol or CE / Lyso PtdCho micelles would offer advantages over existing methods.
Using this technique we demonstrated that such macrophages mimicked biological properties attributed to cholesterol loaded macrophages. Earlier, we had used similar technique to enrich cells with beta carotene. We suggest that this novel technique of delivering macromolecules to the cells could be further manipulated to deliver other hydrophobic large molecular cargos to the cells.