Reduced quenching and extraction time for mammalian cells using filtration and syringe extraction.

ABSTRACT: In order to preserve the in vivo metabolite levels of cells, a quenching protocol must be quickly executed to avoid degradation of labile metabolites either chemically or biologically. In the case of mammalian cell cultures cultivated in complex media, a wash step previous to quenching is necessary to avoid contamination of the cell pellet with extracellular metabolites, which could distort the real intracellular concentration of metabolites. This is typically achieved either by one or multiple centrifugation/wash steps which delay the time until quenching (even harsh centrifugation requires several minutes for processing until the cells are quenched) or filtration. In this article, we describe and evaluate a two-step optimized protocol based on fast filtration by use of a vacuum pump for

SUBMITTER: Hernandez Bort JA 

PROVIDER: S-EPMC4071440 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature