title: Quenching methods for the analysis of intracellular metabolites.
abstract: Sampling and quenching methods for intracellular metabolite analysis in mammalian cells in adherent and suspension culture are described. Quenching of adherent cells is achieved by application of hot air after removal of the supernatant by suction. For suspension cultures, the addition of excess ice-cold saline results in a rapid inactivation of metabolism and significant dilution of extracellular metabolites. Medium carryover is prevented by rinsing the cells with washing solution. Separation of supernatant from suspension cells via centrifugation is incomplete due to required short centrifugation time. Thus, it is necessary to determine the reproducible cell recovery after quenching.
authors.affiliation: Biochemical Engineering, Saarland University, Saarbrücken, Germany.
authors.forename: Judith
authors.initials: J
authors.lastname: Wahrheit
authors.forename: Elmar
authors.initials: E
authors.lastname: Heinzle
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002498:Centrifugation / Q000379:methods; D005109:Extracellular Matrix / Q000378:metabolism; D008322:Mammals / Q000378:metabolism*; D055432:Metabolomics / Q000379:methods*; D013997:Time Factors