Hepatocytes primary culture from the Neotropical fish, trahira Hoplias malabaricus (Bloch)

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Abstract:

Hepatocytes were isolated and cultured from Hoplias malabaricus using a non-enzymatic protocol. From 6·9 to 9·4 × 107 hepatocytes g−1 of liver were isolated and cell viability was 65–85%. Fish hepatocytes attached well to untreated polystyrene flasks (Corning) as well as on treated surfaces with extracellular matrix proteins, i.e. fibronectin, matrigel or type I collagen coatings. Cells rapidly started migrating and reorganizing in cord-like groups after seeding, with spaces similar to bile canaliculi in vivo. Prominent nucleolus and euchromatin-rich rounded nucleus, abundant mitochondria, well-developed rough endoplasmic reticulum, polysomes and Golgi apparatus were seen under ultrastructural analysis. Cells remained functional and metabolically active after 6 days in culture. The protocol established in the current work provides the basis for further studies of native fishes for accurate in vitro toxicological studies.

Keywords: Hoplias malabaricus; hepatocytes; primary cell culture; tropical fishes

Document Type: Regular Paper

DOI: http://dx.doi.org/10.1111/j.1095-8649.2006.01217.x

Affiliations: 1: Departamento de Biologia Celular, Universidade Federal do Paraná, CP 19031, CEP: 81.531-990, Curitiba-PR, Brasil, 2: Departamento de Patologia Básica, Universidade Federal do Paraná, CP 19031, CEP: 81.531-990, Curitiba-PR, Brasil 3: Institut des Sciences de la Mer de Rimouski (ISMER), Université du Québec à Rimouski, 310 allée des Ursulines, Rimouski, Québec, G5L 3A1, Canada

Publication date: November 1, 2006

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