ISOLATION AND PRIMARY CULTURE OF TELEOST HEPATOCYTES

The review discusses procedures for isolation and primary culture of t eleostean hepatocytes and the influence of different culture condition s on the physiology of the cells in vitro. As a routine method to isol ate fish liver cells, enzymatic dissociation of the tissue, either in situ or after removal from the donor animal, is applied. In primary cu lture, piscine hepatocytes usually are maintained as monolayer culture in both serum-free or serum-containing media. Under such simple condi tions, teleostean liver cells conserve viability and functional differ entiation for approximately 5 days. For longer-term culture, in vitro conditions have to be developed which mimic more closely the in vivo s ituation. Medium composition and particularly the cellular micro-envir onment, i.e. the presence of extracellular matrix or of homologous and heterologous cell-cell interactions, appear to be of importance for e xtended conservation of liver-specific gene expression in vitro. Among the various possible alternatives to the monolayer technique: hepatoc yte aggregate cultures are particularly promising. Primary cultures ar e an attractive model to study time-dependent induction processes unde r defined experimental conditions, however, the potential of this syst em to extend our understanding of basic aspects of fish liver physiolo gy and its adaptive responses to environmental change has remained lar gely unexplored. (C) 1998 Elsevier Science Inc. All rights reserved.