無血清培地によるウナギ遊離肝細胞の初代培養〔英文〕

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Primary culture of the hepatocytes of the eel was performed in the serum-free medium. The serum-free medium consisted of Williams'medium E, insulin, glucagon, EGF, prolactin, growth hormone, and H2SeO3. The clear nuclei in the cells cultured in the serum-free medium were observed during 10 to 15days culture and the clear boundaries between the cells as monolayer were also observed. The cells cultured in the medium containing 10% serum accumulated vacuoles like oil droplets and the appearance of nonparenchymal cells was observed after 7 days of culture. The stimulation of DNA synthesis and cell proliferation were recognized in the cells cultured in the serum-free medium. It was found that all hormones and EGF were necessary for the cell proliferation, and that insulin, glucagon, and prolactin were necessary for the maintenance of normal morphology of the cells besides the proliferation. In the primary culture of the hepatocytes of the eel, the use of the dish precoated with fibro-nectin was essential. To investigate the necessity of fibronectin the density of the parenchymal cells of the eel liver was determined by density gradient centrifugation using Percoll. It was found that the density of the parenchymal cells of the eel liver was from 1.025 to 1.045 and lighter than that of rat liver. Therefore, fibronectin seemed to be necessary to avoid floating the cells from the surface of a dish and to attach the cells on the dish.
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