A simple method for isolation and culture of primary hepatocytes from Salvelinus leucomaenis (White-spotted Charr)
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ORIGINAL ARTICLE
A simple method for isolation and culture of primary hepatocytes from Salvelinus leucomaenis (White-spotted Charr) Katsuya Iuchi . Yasushi Arai . Kazuki Sasaki . Naoe Sato . Chikako Yokoyama . Toshiro Saruwatari . Hisashi Hisatomi
Received: 24 March 2020 / Accepted: 4 August 2020 Ó Springer Nature B.V. 2020
Abstract 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 whitespotted charr and used basic methods, such as enzyme digestion and low centrifugation, to analyze the Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10616-020-00415-6) contains supplementary material, which is available to authorized users. K. Iuchi (&) K. Sasaki H. Hisatomi Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, 3-3-1 Kichijojikitamachi, Musashino-shi, Tokyo 180-8633, Japan e-mail: [email protected]; [email protected]
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.
T. Saruwatari Atmosphere and Ocean Research Institute, The Univeristy of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8564, Japan
Y. Arai N. Sato Seikei Junior and Senior High School, 3-10-13 Kichijojikitamachi, Musashino-shi, Tokyo 180-8633, Japan C. Yokoyama Department of Biochemical Engineering, Graduate School of Science and Engineering, Yamagata University, 4-3-16 Jonan, Yonezawa, Yamagata 992-8510, Japan
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Cytotechnology
Keywords Experimental model Cell culture Hepatocytes White-spotted charr
Introduction Salvelinus leucomaenis (S. l.), or white-spotted charr, is an anadromous freshwater fish. The white-spotted charr is distributed from Honshu to Kamchatka Peninsula (Hosoya 2013). The white-spotted charr has been classified into four subspecies (S. l. imbrius, S. l. japonicus, S. l. leucomaenis, and S. l. pluvicus) based on zoogeographic patterns and morphological characteristics (Nakabo 2000). We recently reported the complete mitochondrial DNA sequences of two white-spotted chars (S. l. imbrius, S. l. pluvius) in Japan, as well as those of other charrs (Arai et al. 2019). Recently, th
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