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Hepatocytes primary culture from the Neotropical fish, trahira Hoplias malabaricus (Bloch)
Author(s) -
Filipak Neto F.,
Zanata S. M.,
Randi M. A. F.,
Pelletier É.,
Oliveira Ribeiro C. A.
Publication year - 2006
Publication title -
journal of fish biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.672
H-Index - 115
eISSN - 1095-8649
pISSN - 0022-1112
DOI - 10.1111/j.1095-8649.2006.01217.x
Subject(s) - biology , endoplasmic reticulum , nucleolus , microbiology and biotechnology , golgi apparatus , anatomy , cytoplasm
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.