Use of Primary Cultures of Salmon Hepatocytes for the Study of Hormonal Regulation of Insulin-like Growth Factor I Expression In Vitro

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Hepatocytes isolated from sockeye salmon (Oncorhynchus nerka) were seeded on an uncoated plastic dish with a positively charged surface, and cultured in hormone-free defined medium. Initially single cells began to aggregate and became flat after being attached to the dish. After approximately 6 days in culture, hepatocytes formed a monolayer sheet with cells in side-by-side contact, and remained in this form for at least 2 weeks. Hepatocytes in multicellular spheroids were also observed. The viability of the cells remained high (85-90%) during culture as judged by trypan blue exclusion. The cells actively incorporated [^3H] leucine into cellular and secreted proteins. Salmon and bovine insulins stimulated the leucine incorporation in a dose-dependent manner, suggesting that the cells remained responsive to hormones during culture. The basal IGF-I mRNA level in cultured hepatocytes was much lower than that of in vivo status. Salmon GH increased the level of IGF-I mRNA, whereas salmon prolactin had no effect. These findings suggest that the primary culture system of salmon hepatocytes can be used for the in vitro study of hormonal regulation of IGF-I gene expression in salmon.
社団法人日本動物学会の論文
1993-06-15