Iuchi Katsuya, Arai Yasushi, Sasaki Kazuki, Sato Naoe, Yokoyama Chikako, Saruwatari Toshiro, Hisatomi Hisashi
Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, 3-3-1 Kichijojikitamachi, Musashino-shi, Tokyo, 180-8633, Japan.
Seikei Junior and Senior High School, 3-10-13 Kichijojikitamachi, Musashino-shi, Tokyo, 180-8633, Japan.
Cytotechnology. 2020 Oct;72(5):731-739. doi: 10.1007/s10616-020-00415-6. Epub 2020 Aug 10.
White-spotted charr (Salvelinus leucomaenis, S. I.) is an anadromous cold water-adapted fish, distributed in the Far East. We have previously reported the complete mitochondrial DNA sequences of white-spotted chars (S. l. imbrius and S. l. pluvius) in Japan. In general, fish hepatocytes are useful for cellular and biochemical studies of fish. In this study, we isolated hepatocytes from the liver of white-spotted charr and used basic methods, such as enzyme digestion and low centrifugation, to analyze the molecular mechanisms involved in specific cellular responses. The isolated hepatocytes could be cultured at 5-20 °C but not 37 °C. The morphology of hepatocytes was altered in a temperature-dependent manner. The properties of hepatocyte were similar to those of living fish. Moreover, the proliferation rate and damage of isolated hepatocytes depended on the concentration of fetal bovine serum in the culture medium. Taken together, this study demonstrates that this simple method for isolation and culture of hepatocytes from white-spotted charr may be useful for other biochemical and cellular studies.
白斑红点鲑（Salvelinus leucomaenis，S. I.）是一种适应溯河洄游的冷水性鱼类，分布于远东地区。我们之前已经报道了日本白斑红点鲑（S. l. imbrius和S. l. pluvius）的线粒体DNA全序列。一般来说，鱼类肝细胞对于鱼类的细胞和生化研究很有用。在本研究中，我们从白斑红点鲑的肝脏中分离出肝细胞，并使用酶消化和低速离心等基本方法来分析特定细胞反应所涉及的分子机制。分离出的肝细胞可以在5 - 20°C下培养，但不能在37°C下培养。肝细胞的形态以温度依赖的方式发生改变。肝细胞的特性与活鱼相似。此外，分离出的肝细胞的增殖率和损伤程度取决于培养基中胎牛血清的浓度。综上所述，本研究表明这种从白斑红点鲑中分离和培养肝细胞的简单方法可能对其他生化和细胞研究有用。
