Characterization of Non-Cholesterol Sterols in Microglia Cell Membranes Using Targeted Mass Spectrometry

ABSTRACT: Background: Non-cholesterol sterols, as well as plant sterols, cross the blood–brain barrier and, thus, can be incorporated into cell membranes, affecting the cell’s inflammatory response. The aim of our work was to develop an analytical protocol for a quantitative assessment of the sterol composition within the membrane microdomains of microglia. Methods: A protocol for cell membrane isolation using OptiPrepTM gradient ultracentrifugation, in combination with a targeted mass spectrometry (LC-MS/MS)-based assay, was developed and validated for the quantitative analysis of free sterols in microglia cell membranes. Results: Utilizing an established LC-MS/MS assay, cholesterol and seven non-cholesterol sterols were analyzed with a limit of detection from 0.001 to 0.05 mg/L. Applying the deter

SUBMITTER: Begcevic Brkovic I 

PROVIDER: S-EPMC10093698 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature