Grandl, Margot and Schmitz, Gerd (2010) Fluorescent High-Content Imaging Allows the Discrimination and Quantitation of E-LDL-Induced Lipid Droplets and Ox-LDL-Generated Phospholipidosis in Human Macrophages. CYTOMETRY PART A, 77A (3). pp. 231-242. ISSN 1552-4922, 1552-4930
Full text not available from this repository. (Request a copy)Abstract
Macrophage foam cells formed during uptake of atherogenic lipoproteins are a hallmark of atherosclerotic lesion development. In this study, human macrophages were incubated with two prototypic atherogenic LDL modifications enzymatically degraded LDL (E-LDL) and oxidized LDL (Ox-LDL) prepared from the same donor LDL. To detect differences in macrophage lipid storage, fluorescent high-content imaging was used. Lipid droplets were stained using Bodipy 493/503, and the fluorescent phospholipid probe NBD-PE was used to detect endolysosomal phospholipidosis in high-content imaging assays. The phospholipidosis assay was validated using phospholipidosis-inducing cationic amphiphilic drugs. In addition, neutral lipids and phospholipidosis were determined using LipidTOX. Images of 96-well cell culture microtiter plates were captured with multichannel laser-based high-content confocal microscopy, and subsequently cell- and well-based data were analyzed. E-LDL-loaded macrophages show increased intensity of Bodipy 493/503 and LipidTOX(TM)-Green neutral lipid droplet staining and a greater mean area and number of lipid droplets per cell compared to Ox-LDL-loaded and M-CSF-differentiated control macrophages. In contrast, Ox-LDL-loaded macrophages show increased intensity of NBD-PE and LipidTOX(TM)-Red detectable phospholipidosis in the endolysosomal compartment compared to E-LDL-loaded and M-CSF-differentiated macrophages. Treatment with the peroxisome proliferator-activated receptor-gamma agonist pioglitazone leads to lipid droplet induction depending on the lipid loading state of the macrophages. These results indicate that E-LDL preferentially induces lipid droplets, while Ox-LDL provokes endolysosomal phospholipidosis in human macrophages representing two different lipid storage principles. Therefore, fluorescent high-content imaging is a useful tool to discriminate between and quantify lipid storage compartments in macrophages also in response to drugs affecting cellular lipid metabolism. (C) 2009 International Society for Advancement of Cytometry
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | LOW-DENSITY-LIPOPROTEIN; DRUG-INDUCED PHOSPHOLIPIDOSIS; IN-VITRO; NILE RED; ATHEROSCLEROTIC LESIONS; GENE-EXPRESSION; UP-REGULATION; OXIDIZED LDL; CELLS; CHOLESTEROL; E-LDL; Ox-LDL; human macrophages; lipid droplets; phospholipidosis |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 05 Aug 2020 04:50 |
| Last Modified: | 05 Aug 2020 04:50 |
| URI: | https://pred.uni-regensburg.de/id/eprint/25022 |
Actions (login required)
 |
View Item |
