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A dual enzyme method for the established of long-and medium-term primary cultures of epithelial and fibroblastic cells for Atlantic salmon gills

Citation

Butler, R and Nowak, BF, A dual enzyme method for the established of long-and medium-term primary cultures of epithelial and fibroblastic cells for Atlantic salmon gills, Journal of Fish Biology, 65, (4) pp. 1108-1125. ISSN 0022-1112 (2004) [Refereed Article]

DOI: doi:10.1111/j.1095-8649.2004.00521.x

Abstract

A dual enzyme disaggregation method using collagenase and then trypsin was developed that allowed the reproducible initiation of primary cultures from Atlantic salmon Salmo salar gills. Cultures had both epithelial and fibroblast morphology and persisted for an average of 20 passages. Growth was dependent upon a minimum concentration of 5% foetal calf serum (FCS) for fibroblasts and 10% FCS for epithelial cells. Growth was mostly independent of substrate, although epithelial cells showed increased growth on type 1 collagen gels. Matrigel™ cell culture substrate produced reduced growth of fibroblasts and did not benefit epithelial cell growth. Epithelial cells reacted with monoclonal antibodies (MAbs) against mammalian cytokeratins, and fibroblast cells reacted with MAbs against mammalian fibronectin and type 1 collagen. The method also produced two long-term cultures: one epithelial and one fibroblast that have been designated RGE-2 and RGF respectively. © 2004 The Fisheries Society of the British Isles.

Item Details

Item Type:Refereed Article
Research Division:Agricultural and Veterinary Sciences
Research Group:Fisheries Sciences
Research Field:Fish Pests and Diseases
Objective Division:Animal Production and Animal Primary Products
Objective Group:Fisheries - Aquaculture
Objective Field:Fisheries - Aquaculture not elsewhere classified
Author:Butler, R (Dr Rick Butler)
Author:Nowak, BF (Professor Barbara Nowak)
ID Code:30051
Year Published:2004
Web of Science® Times Cited:5
Deposited By:TAFI - Aquaculture
Deposited On:2004-08-01
Last Modified:2005-05-02
Downloads:0

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