17 Toda Robert, Aiko McQuade, Amanda Koppes-den Hertog, Sascha J Erlebach, Lena Kronenberg-Versteeg, Deborah Kampmann, Martin Giera, Martin van der Kant, Rik AG Jucker AG Kronenberg-Versteeg Comparative lipidomics of iPSC-derived microglia protocols reveal lipid droplet and immune differences mediated by media composition. Stem cell reports Stem cell reports Cell Press Maryland Heights, MO 2213-6711 10.1016/j.stemcr.2025.102779 English 102779 2 21 Altered microglial lipid metabolism is heavily implicated in Alzheimer's disease (AD) and aging. Recently, protocols were developed to generate human induced pluripotent stem cell-derived microglia-like cells (iMGL) to study microglial function in vitro, including embryoid body-based methods and induced transcription factor (iTF)-dependent approaches. Here, we performed comparative lipidomics on iMGL from these methods and report major differences in multiple lipid classes, including triglycerides (TGs), a storage form of fatty acids implicated in microglial reactivity. TGs are strongly increased in iTF microglia due to the absence of a media supplement (B-27). Supplementing iTF microglia with B-27, or its component L-carnitine, reduces TGs and promotes a homeostatic state. B-27 also renders iTF microglia metabolically responsive to immune stimuli. Overall, our data show that iMGL differentiation methods have a major impact on microglial lipidomes and warrant attention when studying AD and neuroinflammatory processes involving lipids. Microglia: metabolism Microglia: cytology Microglia: drug effects Microglia: immunology Induced Pluripotent Stem Cells: cytology Induced Pluripotent Stem Cells: metabolism Induced Pluripotent Stem Cells: drug effects Humans Lipidomics: methods Lipid Droplets: metabolism Culture Media: chemistry Culture Media: pharmacology Cell Differentiation: drug effects Lipid Metabolism Triglycerides: metabolism Cells, Cultured iPSC lipid droplet lipid metabolism lipidomics microglia neuroinflammation triglycerides Culture Media Triglycerides Journal Article 2026 DZNE-2026-00189 2026 pmid:41512865 https://pub.dzne.de/record/285210 https://doi.org/10.1016/j.stemcr.2025.102779