Fast filtration for metabolome sampling of suspended animal cells

Compartment‐specific metabolomics for CHO reveals that ATP pools in mitochondria are much lower than in cytosol

“…ATP-pools in CHO cells have been reported to be in the range of 1.5-4.7 fmol cell -1 [15,18]. We found initial pool sizes about 3 fmol cell -1 which fit fairly well to the other studies.…”

“…The corresponding pool sizes in our study were 0.04, 0.1 and 0.05 fmol cell -1 , respectively. Summarizing, considering the different cell types and cultivation conditions applied intracellular pool sizes of this study fit fairly well to independent measurements of others [15,18,47].…”

“…We found initial pool sizes about 3 fmol cell -1 which fit fairly well to the other studies. Furthermore pyruvate was measured at 6 fmol cell -1 [15], while the initial values of our studies revealed 2.5 fmol cell -1 . Intracellular pool sizes of 0.09, 0.89 and 0.03 fmol cell -1 for 3PG, citrate and G6P in MDCK cells, as well as 0.13, 2.05 and 0.23 fmol cell -1 in Vero cells were presented by Ritter et al [47].…”

“…Volmer et al [15] had already succeeded in implementing a related filtering method. However, as our approach applied different filter and housing materials this issue was re-investigated.…”

“…Whole cells are quenched in ice-cold methanol [13], spun down in ice-cold phosphate buffer or saline solution [14], simultaneously filtered and quenched [15] or exposed to heat [16]. In addition to the well-known problems of leakage [17,18], temperature induced degradation of metabolites or interactions with filter materials [19], these protocols only yield averaged, cellular metabolite levels.…”