Three-dimensional Cell Culture Systems Stabilise the Differentiation of Hepatocyte Cell Lines

Abstract

The suitability of a hepatocyte cell line for in vitro toxicity and drug metabolism largely depends in the expression of hepatic differentiated functions. In the present study the activity of phase I enzymes was examined in a long-term run using immobilised (Cultispher GH) and aggregates of hepatocyte cell lines derived from Sprague-Dawley male rats. With that purpose the metabolic profile of testosterone was followed as the oxidative metabolism of this hormone is catalysed, with a high degree of regio- and stereoselectivity, by different cytochrome P450 families (1 A, 2A, 2B, 2C and 3A). The results revealed that the cytochrome P450 activities were maintained for higher periods of time in three-dimensional cell systems then monolayer (conventional) cell cultures. Conventional cultures showed a rapid loss of cytochrome P450, which were undetectable after few passages. In contrast, some isoenzymes assayed were measurable in three-dimensional cell cultures systems for as long as 4 months. The beneficial effects of three-dimensional systems seemed to be attributed to the cell-cell contacts that will promote their phenotypic stability or to the retention of globular cell structure.