title: Fast filtration for metabolome sampling of suspended animal cells.
abstract: A new method for sampling suspended animal cells by fast filtration is presented that allows rapid quenching of cellular metabolism and efficient separation of the cells from culture medium. Compared to sampling with a microstructure heat exchanger or centrifugation without prior quenching, the adenylate energy charge and the measured concentrations especially of metabolites with a high turnover rate or of metabolites early in metabolic pathways were substantially higher. No leakage of ATP from the cells was observed when using iso-osmotic NaCl solution in the washing step. The combination of fast filtration and cold methanol extraction is therefore suitable for intracellular metabolomic studies of suspended animal cell cultures and superior to other methods currently applied.
authors.affiliation: Institute of Cell Culture Technology, Bielefeld University, Bielefeld, Germany. martin.volmer@uni-bielefeld.de
authors.forename: Martin
authors.initials: M
authors.lastname: Volmer
authors.forename: Stefan
authors.initials: S
authors.lastname: Northoff
authors.forename: Sebastian
authors.initials: S
authors.lastname: Scholz
authors.forename: Tobias
authors.initials: T
authors.lastname: Thüte
authors.forename: Heino
authors.initials: H
authors.lastname: Büntemeyer
authors.forename: Thomas
authors.initials: T
authors.lastname: Noll
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005374:Filtration / Q000379:methods*; D055432:Metabolomics