Comparison of two cell isolation procedures to study in vitro intestinal wall biotransformation in control and 3‐methyl‐cholanthrene pretreated rats

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Abstract
Two cell isolation procedures, i.e. a scraping/collagenase‐treatment and a new vibration procedure in EDTA containing medium, were used to isolate intestinal epithelial cells. In both cell populations the metabolism of 7‐ethoxycoumarin and 7‐hydroxycoumarin was studied. Moreover, the time course and extent of induction of both steps in the biotransformation were investigated after oral 3‐methylcholanthrene pretreatment of the rats. Twenty four hours after 3‐methylcholanthrene pretreatment (20 mg kg−1) monooxygenase activity was induced about 6‐fold and 2.5‐fold when studied with cells of the vibratory and enzymic procedures, respectively. Control 7‐ethoxycoumarin deethylase activity and 7‐hydroxycoumarin glucuronidation were about the same when comparing both methods for cell‐isolation. The formation of glucuronides in cells (both methods) is significantly lowered by 3‐MC pretreatment, while sulphation remains unaffected. Results indicate that using enzymic treatment of mucosal scrapings, cell‐populations are obtained containing relatively more differentiated (tip) cells. A number of advantages of the new (vibration) method are: better recovery, viability and reproducibility.