Abstract
Hepatocytes were isolated and cultured from Hoplias malabaricus using a non‐enzymatic protocol. From 6·9 to 9·4 × 10 7 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.