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The influence of the choice of digestion enzyme used to prepare rat hepatocytes on xenobiotic uptake and efflux

Sinclair JA, Henderson C, Tettey JN, Grant MH

Toxicology in vitro : an international journal published in association with BIBRA, 2013

Abstract

Isolated rat hepatocytes are widely used to assess the metabolism and toxicity of xenobiotics. The choice of digestion enzyme used to prepare the cells has been shown previously to influence their metabolic capability. This study investigates the effect of the digestion enzyme (collagenase II, collagenase A/trypsin inhibitor, or collagenase plus dispase) on the uptake of xenobiotics into, and efflux from, hepatocytes. The choice of digestion enzymes used in this study does not affect uptake of either pravastatin (an organic anion probe substrate for Oatp transporter) or metformin (an organic cation probe substrate for Oct transporter). With regard to efflux transporters, hepatocyte differentiation was better maintained when cells were isolated using collagenase II alone.

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