Calf primary hepatocyte culture as a tool for anabolic steroid metabolism studies†

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Abstract
Calf hepatocyte cultures were developed for the predictive analysis of in vivo xenobiotic biotransformations. The goal of this work was to show the feasibility of a system based on calf primary hepatocyte cultures using radiolabelled molecules to study xenobiotic metabolism rather than to study exhaustively the metabolism of a particular steroid. 19-Nortestosterone was chosen as a model substrate because of its relatively well known metabolism in the bovine species. Incubation of a mixture of tritiated and non-tritiated 19-nortestosterone was investigated in cell culture in order to target the biosynthesized metabolites. After extraction and purification, some of the in vitro metabolite structures were elucidated by GC-MS and compared with the main urinary metabolites detected in vivo after intramuscular injection of 19-nortestosterone into a calf. 19-Norepitestosterone, the main in vivo metabolite, was identified in vitro. However, the main in vitro metabolites were mostly in an oxidized form (4-estrene-3,17-dione, hydroxy-4-estrene-3,17-dione).

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