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Procedures for the reconstruction, primary culture and experimental use of rainbow trout gill epithelia

Research output: Contribution to journalArticle

Sabine Schnell, Lucy Claire Stott, Christer Hogstrand, Chris Wood, Scott Kelly, Peter Part, Stewart Owen, Nicolas Richard Bury

Original languageEnglish
Pages (from-to)490-498
Number of pages9
JournalNature Protocols
Volume11
Issue number3
DOIs
Publication statusE-pub ahead of print - 11 Feb 2016

Documents

  • NP-PI150167B Schnell et al

    NP_PI150167B_Schnell_et_al.docx, 6.29 MB, application/vnd.openxmlformats-officedocument.wordprocessingml.document

    11/08/2016

    Accepted author manuscript

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King's Authors

Abstract

This protocol describes how to reconstruct and culture the freshwater rainbow trout gill epithelium on flat permeable membrane supports within cell culture inserts. The protocol describes gill cell isolation, cultured gill epithelium formation, maintenance, monitoring and preparation for use in experimental procedures. To produce a heterogeneous gill epithelium, as seen in vivo, seeding of isolated gill cells twice over a two day period is required. As a consequence this is termed the double seeded insert (DSI) technique. Approximately 5-12 days after cell isolation and seeding, preparations develop electrically tight gill epithelia that can withstand fresh water on the apical cell surface. The system can be used to study freshwater gill physiology, as well as a humane alternative for toxicity testing, bioaccumulation studies and environmental water quality monitoring.

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