Isolation and primary culture of teleost hepatocytes

Abstract

The review discusses procedures for isolation and primary culture of teleostean hepatocytes and the influence of different culture conditions on the physiology of the cells in vitro. As a routine method to isolate fish liver cells, enzymatic dissociation of the tissue, either in situ or after removal from the donor animal, is applied. In primary culture, piscine hepatocytes usually are maintained as monolayer culture in both serum-free or serum-containing media. Under such simple conditions, teleostean liver cells conserve viability and functional differentiation for approximately 5 days. For longer-term culture, in vitro conditions have to be developed which mimic more closely the in vivo situation. Medium composition and particularly the cellular micro-environment, i.e. the presence of extracellular matrix or of homologous and heterologous cell–cell interactions, appear to be of importance for extended conservation of liver-specific gene expression in vitro. Among the various possible alternatives to the monolayer technique, hepatocyte aggregate cultures are particularly promising. Primary cultures are an attractive model to study time-dependent induction processes under defined experimental conditions, however, the potential of this system to extend our understanding of basic aspects of fish liver physiology and its adaptive responses to environmental change has remained largely unexplored.