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

Schnell, S, Stott, L.C, Hogstrand, C, Wood, C.M, Kelly, S.P, Part, P, Owen, S.F and Bury, Nic (2016) Procedures for the reconstruction, primary culture and experimental use of rainbow trout epithelia. Nature Protocols, 11 (3). pp. 490-498. ISSN 1754-2189

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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 2-d period is required. As a consequence, this is termed the double-seeded insert technique. Approximately 5-12 d after cell isolation and seeding, preparations develop electrically tight gill epithelia that can withstand freshwater on the apical cell surface. The system can be used to study freshwater gill physiology, and it is a humane alternative for toxicity testing, bioaccumulation studies and environmental water quality monitoring.

Item Type: Article
Uncontrolled Keywords: Cell culture, Ecotoxicology, Environmental monitoring, Non-model organisms
Subjects: Q Science > Q Science (General)
Q Science > QH Natural history
Divisions: Faculty of Health & Science > Department of Science & Technology
Depositing User: David Upson-Dale
Date Deposited: 27 Feb 2017 11:52
Last Modified: 23 Nov 2017 09:35
URI: http://oars.uos.ac.uk/id/eprint/188

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