Viability and induction of tyrosine aminotransferase in rainbow trout hepatocytes cultured on laminin and polylysine in a serum-free medium

Abstract

The objective of this study was to find an economical substrate for long-term primary culture of rainbow trout hepatocytes in which viability and differentiated liver function could be maintained in a serum-free media. Attachment efficiency of primary cultures of rainbow trout (Oncorhynchus mykiss) hepatocytes was determined for cells grown on plastic and Falcon Primaria dishes, and dishes coated with ECM (extracellular matrix), Matrigel, fibronectin, collagen type 1, laminin and polylysine. On ECM, Matrigel, laminin and polylysine, attachment efficiency exceeded 90%. On plastic, Falcon Primaria, fibronectin and collagen-coated dishes attachment efficiency was 15–20% and the cells were easily dislodged by pipetting. Cells grown on ECM, laminin and polylysine were maintained up to 22 day with 76%, 60% and 80% viability, respectively. On ECM cells flattened and formed a confluent monolayer on day 2 in culture. Cells grown on matrigel, laminin and polylysine had a more rounded appearance with establishment of cell-to-cell contacts on the second day in culture. Induction of TAT (tyrosine aminotransferase), a marker enzyme for differentiated liver cell functions, was studied with DEX (dexamethasone) in cells grown on laminin and polylysine in serum-free media. Cells grown on laminin and polylysine showed TAT inducibility up to 15 and 19 days, respectively.