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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 
 
 

Phase I and II biotransformations in living CaCo 2 cells cultivated under serum-free conditions. Selective apical excretion of reaction products.

CaCo 2 cells, cultivated in a synthetic, serum-free nutritive medium on poly (ethylene terephthalate) membranes, form a confluent monolayer of differentiated cells, with the apical and basolateral poles exposed to the upper and lower compartments, respectively, of bicameral culture inserts (Halleux and Schneider, In Vitro Cell Dev Biol, 27A: 293-302, 1991). This cell culture system allows the passage of intact mannitol by the paracellular route and the transcellular diffusion of testosterone which appears mainly as a biotransformed unconjugated metabolite. When ethoxyresorufin is added to either the apical or basolateral poles of living CaCo 2 cells, resorufin is formed, and more than 80% is excreted at the apical pole. Under our experimental conditions, no detectable amounts of glucurono- or sulfconjugates are found. Methylcholanthrene and phenobarbital increase the biotransformation of ethoxyresorufin 50 and 3 times, respectively, and induce that of benzoxyresorufin, but not of pentoxyresorufin which remains absent under all conditions. These substances do not affect the proportion of resorufin recovered at the apical role. Verapamil inhibits by 25% the release of resorufin but does not affect its distribution. Chlorodinitrobenzene is conjugated with glutathione and at least two-thirds of the product is excreted at the apical pole; methylcholanthrene and phenobarbital do not increase this activity. These results demonstrate that differentiated CaCo 2 cells, under serum-free conditions, perform phase I and II reactions and that the biotransformation products are selectively excreted at the apical pole.[1]

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