Heterogeneity and standardization of phase II metabolism in cultured cells.
Caco-2 cells are widely used for transepithelial transport and metabolism studies. We analysed the metabolites produced from quercetin (Q) during transport of this flavonoid across Caco-2 monolayers and by plastic-adhering cells. We found that the pattern of Phase II metabolic activity varies markedly depending on the particular cell clone, age of the cell culture, and stressful treatment such as freezing/thawing. Prolonged culturing and stress cause a decrease of "detoxifying" conjugating activity. This can be re-established by growing the cells with a low concentration of the transport/metabolism substrate for a few days. We suggest this metabolism-activating procedure be used to make studies with these cells more readily comparable.
Biasutto
Lucia
L
Department of Biomedical Sciences, University of Padova, Italy.
Marotta
Ester
E
De Marchi
Umberto
U
Beltramello
Silvia
S
Bradaschia
Alice
A
Garbisa
Spiridione
S
Zoratti
Mario
M
Paradisi
Cristina
C
D018938:Caco-2 Cells; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D005615:Freezing; D006801:Humans; D050217:Metabolic Detoxication, Phase II; D011794:Quercetin / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D015238:Sulfotransferases / Q000378:metabolism
D011794:Quercetin; D015238:Sulfotransferases