A new isolating procedure of human hepatocytes has been developed using two-step collagenase digestion by a nonperfusion procedure (NP) of non-wedge liver biopsies.
1. A yield of 2 - 76106 hepatocytes/g liver, 52 - 95% viability and 13 - 75% attachment were obtained from liver biopsies weighing 6 - 60 g, comparable to that obtained when using the classical perfusion procedure (P) to isolate human hepatocytes from wedge liver samples of 50 - 150 g.
2. In culture, human hepatocytes obtained by NP remained attached to plastic for up to 5 days and displayed the usual morphological characteristics.
Their metabolic capacities, assessed by liver-specific albumin and urea synthesis and by CYP-dependent and conjugation pathways, were equivalent to those of human hepatocytes obtained by P. In addition, they responded adequately to specific CYP inducers, demonstrating that they constitute a model in which human drug metabolism and toxicity studies can be performed.
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