Optimizing methods of measuring metabolic flux using liquid chromatography-mass spectrometry

Department

North Carolina Agricultural and Technical State University

Document Type

Poster

Publication Date

4-17-2026

Abstract

Organoids are three-dimensional, immortalized human cell cultures used in biomedical research to study cell physiology and disease progression. Analyzing how these cells metabolize nutrients is essential to understanding how cells function and how disruptions to metabolism contribute to disease. However, conducting such an analysis on whole organoids with multiple cell types yields convoluted results as metabolic fluxes are not distinguishable between cell types. Sorting organoid cells before metabolomic analysis reveals the unique metabolism of each cell type, but the separation process risks altering metabolic fluxes before analysis. Our project aims to optimize methods for separating, quenching, and analyzing metabolism in individual cell types within brain organoids using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Currently, we have developed a metabolite library of citric acid cycle intermediates, including their MS/MS transition patterns. Currently, we are using our method to analyze the metabolites within single-cell extracts under various conditions. In the future, we will experiment with assembling and separating organoids with magnetic-assisted cell separation (MACS). Once developed, this method opens the door to new experiments to understand the complex physiology of the brain and other organs in disease states via LC-MS/MS-based metabolomics.

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