Faculty, Staff and Student Publications

Language

English

Publication Date

2-1-2026

Journal

Life Science Alliance

DOI

10.26508/lsa.202503529

PMID

41371957

PMCID

PMC12696398

PubMedCentral® Posted Date

12-10-2025

PubMedCentral® Full Text Version

Post-print

Abstract

The preparation of custom-made media offers precise control over nutrient composition, enabling detailed studies of cellular metabolism. We demonstrate how self-made media formulations enable diverse assay designs and readouts to assess cancer metabolism. Self-made media can be used in Seahorse assays to measure mitochondrial respiration under defined conditions. In nutrient deprivation experiments, amino acid or vitamin removal can uncover how cancer cells adapt to metabolic stress. Using labeled amino acids enables analysis of nascent protein synthesis and translational regulation, while stable-isotope tracing reveals metabolic fluxes through key pathways. This guide presents a suite of metabolic assays using custom-made media, covering experimental design, the selection of controls, sample preparation, data acquisition, and interpretation. The accompanying online media calculator "Media Minds" streamlines the creation of custom media formulations, ensuring accuracy and reproducibility.

Keywords

Humans, Neoplasms, Culture Media, Mitochondria, Cell Line, Tumor, Amino Acids, Metabolomics

Published Open-Access

yes

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