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 of ex vivo cultures 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 this in vitro system 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.

New Stable Cell Lines Derived from the Proximal and Distal Intestine of Rainbow Trout (Oncorhynchus mykiss) Retain Several Properties Observed In Vivo

“…Although primary cell cultures from the different parts of the RT intestinal tract have been derived before, these cells had a finite lifespan of about 6 weeks [ 24 ]. Therefore, to the best of our knowledge, this makes RTpi-MI the first stable cell line derived from the proximal intestine of rainbow trout.…”

Organoids: a promising new in vitro platform in livestock and veterinary research

“…This also applies to other farmed animal species not covered in this review, such as rabbit [ 104 ] and fish species, e.g. rainbow trout [ 105 ]. However, to increase the scope of (large scale) application of organoids several challenges need to be addressed.…”

Immune Transcriptional Response in Head Kidney Primary Cell Cultures Isolated from the Three Most Important Species in Chilean Salmonids Aquaculture

“…Innate immunity has an essential role in protecting fish from endogenous and exogenous pathogenic invasion. Meanwhile, adaptive immunity is a more specific barrier [18,32]. For this reason, to have primary cell culture as the standard method to reduce the number of fish employed in research is very important, including for animal welfare.…”

“…Cell culture is highly relevant as it minimizes animal usage and has been instrumental in studying viral and microbial pathogens in humans and animals [26,27]. An increasing number of specific in vitro methods have been developed for fish [28][29][30], including primary fish gill and liver culture [31] and intestinal epithelial cells [32]. These can be initiated from explants or enzymatic dissociation, require less adaptation to culture media, are economical and represent more closely what physiologically occurs in the host [29,30], allowing the study of more specific responses between species as in this work.…”