Quantitative analysis of red blood cell membrane phospholipids and modulation of cell-macrophage interactions using cyclodextrins

The plasma membrane of eukaryotic cells is asymmetric with respect to its phospholipid composition. Analysis of the lipid composition of the outer leaflet is important for understanding cell membrane biology in health and disease. Here, a method based on cyclodextrin-mediated lipid exchange to chara...

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Bibliographic Details
Published in:Scientific reports 2020-09, Vol.10 (1), p.15111-15111, Article 15111
Main Authors: Vahedi, Amid, Bigdelou, Parnian, Farnoud, Amir M.
Format: Article
Language:English
Subjects:
631/45/287
631/45/287/1192
631/45/608
631/57/2271
Cell Communication
Cyclodextrins - metabolism
Erythrocyte Membrane - metabolism
Erythrocytes - metabolism
Humanities and Social Sciences
Humans
Lipid Bilayers - metabolism
Macrophages - metabolism
Membrane Lipids - metabolism
multidisciplinary
Phospholipids - analysis
Science
Science (multidisciplinary)
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Summary:The plasma membrane of eukaryotic cells is asymmetric with respect to its phospholipid composition. Analysis of the lipid composition of the outer leaflet is important for understanding cell membrane biology in health and disease. Here, a method based on cyclodextrin-mediated lipid exchange to characterize the phospholipids in the outer leaflet of red blood cells (RBCs) is reported. Methyl-α-cyclodextrin, loaded with exogenous lipids, was used to extract phospholipids from the membrane outer leaflet, while delivering lipids to the cell to maintain cell membrane integrity. Thin layer chromatography and lipidomics demonstrated that the extracted lipids were from the membrane outer leaflet. Phosphatidylcholines (PC) and sphingomyelins (SM) were the most abundant phospholipids in the RBCs outer leaflet with PC 34:1 and SM 34:1 being the most abundant species. Fluorescence quenching confirmed the delivery of exogenous lipids to the cell outer leaflet. The developed lipid exchange method was then used to remove phosphatidylserine, a phagocyte recognition marker, from the outer leaflet of senescent RBCs. Senescent RBCs with reconstituted membranes were phagocytosed in significantly lower amounts compared to control cells, demonstrating the efficiency of the lipid exchange process and its application in modifying cell–cell interactions.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-72176-3