Comparison of two cell isolation procedures to study in vitro intestinal wall biotransformation in control and 3-methyl-cholanthrene pretreated rats.
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.
D000818:Animals; D001711:Biotransformation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D003374:Coumarins / Q000378:metabolism*; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism*; D008297:Male; D008748:Methylcholanthrene / Q000494:pharmacology*; D051381:Rats; D014468:Umbelliferones / Q000378:metabolism*
D003374:Coumarins; D014468:Umbelliferones; C017299:7-ethoxycoumarin; D008748:Methylcholanthrene; C031477:7-hydroxycoumarin