Establishment of fish hepatocyte cultures for use in in vitro carcinogenicity studies
Klaunig, J.E.
National Cancer Institute Monograph 65: 163-173
1984
ISSN/ISBN: 0083-1921 PMID: 6431289 Document Number: 227679
Methods were developed for the isolation and primary culture of rainbow trout (Salmo gairdneri) and channel catfish (Ictalurus punctatus) liver cells. Using a two-step perfusion technique, I isolated an average of 2.75 and 2.87 liver cells/g body weight from the trout and catfish, respectively. Hepatocytes represented 91.4% (trout) and 90.1% (catfish) of the total liver cells isolated. Both catfish and trout hepatocytes in primary culture displayed a linear decrease in survival with increased duration of culture. The DNA synthesis in the hepatocytes during culture showed a similar decrease with increased time in culture. Approximately 2.8% (trout) and 3.5% (catfish) of the hepatocytes exhibited nuclear labeling with pyrene, and N-methyl-N'-nitro-N-nitrosoguanidine. Trout hepatocytes displayed a decrease in UDS induction with aflatoxin B1 with increased age of the cultures. However, UDS induced by N-methyl-N'-nitro-N-nitrosoguanidine remained constant throughout the culture period.