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GBM Cell Lines Reproducibility Pilot Study

ABSTRACT: GBM cell lines were plated onto 10 cm dishes at 100,000-200,000 cells per dish and allowed to grow for 3-4 days until confluency reached 50-70%. Media for all cell lines was DMEM with 10% FBS including 25 mM glucose, 6.2 mM glutamine and 200 uM Oleic Acid (conjugated to BSA). 1-2 hours prior to tracer incubation, media was aspirated and fresh media was used. Immediately prior to tracer incubation, media was again aspirated and then cells were washed with warm PBS to remove unlabeled metabolites. Cells were then incubated with labeled tracer (DMEM with 10% FBS including 25 mM Uniformly labeled 13C-6 glucose, 6.2 mM unlabeled glutamine and 200 uM unlabeled oleic acid). After 2 hours of incubation with tracer, media was aspirated, cells were quickly washed with 15 mL DI water, quenched with liquid nitrogen and then stored at -80oC until analysis.

ORGANISM(S): Human Homo Sapiens

TISSUE(S): Glioma Cells

SUBMITTER: Maureen Kachman

PROVIDER: ST000301 | MetabolomicsWorkbench | Tue Mar 17 00:00:00 GMT 2015

REPOSITORIES: MetabolomicsWorkbench

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