Establishment and long-term maintenance of primary intestinal epithelial cells cultured from the rainbow trout,Oncorhynchus mykiss.

Abstract

A novel method for the establishment and long-term maintenance ofex vivocultures from intestinal regions of the rainbow trout,Oncorhynchus mykiss(Walbaum), is reported. Adherence of cells was observed within hours, epithelial island formation recorded at 48 h and rapid proliferation with confluence achieved between 9-14 days. In addition to metabolic characterisation, basic morphology of growing cells was characterised using histology, immunofluorescence, transmission electron microscopy (TEM) and transepithelial electrical resistance (TEER). Regional differences in intestinal ethoxyresorufin-O-deethylase (EROD) and 7-ethoxycoumarin-O-deethylation (ECOD) activities in these primary grown enterocytes were compared following exposure to model inducers [i.e.α-NF,β-NF, B(a)P] which demonstrated significant differences. Regional differences in dietary uptake and metabolism of contaminants can therefore be studied in thisin vitrosystem to increase our understanding of fundamental processes, while concurrently providing a means to reduce the number of fish required for biological studies in line with the principles of the 3Rs (Reduce, Refine and Replace).This article has an associated First Person interview with the first author of the paper.