Abstract
This protocol describes the culturing of the nematode Caenorhabditis elegans (C. elegans) in a sterol-defined experimental system and the subsequent quantitative analysis of C. elegans sterols through gas chromatography-mass spectrometry. Although studied primarily in mammals, sterols are essential biomolecules for most eukaryotes. C. elegans cannot synthesize sterols and thus relies on the uptake of dietary sterols. Therefore, C. elegans is a powerful system to study metabolism in sterol-defined conditions that are described in our protocol. For complete details on the use and execution of this protocol, please refer to Shamsuzzama et al. (2020).