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建立易危鱼类马雷纳白鲑(Coregonus maraena)的体外模型:生长条件的优化及细胞系的特性分析

Establishment of an in vitro model from the vulnerable fish species Coregonus maraena (maraena whitefish): Optimization of growth conditions and characterization of the cell line.

作者信息

Kaya Yagmur, Tönißen Katrin, Verleih Marieke, Rebl Henrike, Grunow Bianka

机构信息

Research Institute for Farm Animal Biology (FBN), Institute of Muscle Biology and Growth, Dummerstorf, Germany.

Research Institute for Farm Animal Biology (FBN), Institute of Genome Biology, Dummerstorf, Germany.

出版信息

Cell Biol Int. 2023 Mar;47(3):548-559. doi: 10.1002/cbin.11956. Epub 2022 Nov 9.

Abstract

In this study, a cell line of the fish species Coregonus maraena was produced for the first time. C. maraena is an endangered species, and studies indicate that this fish species will be affected by further population declines due to climate change. This cell line, designated CMAfin1, has been maintained in Leibovitz L-15 supplemented with 10% fetal bovine serum over 3 years. Both subculturing and storage (short-term storage at -80°C and long-term storage in liquid nitrogen) was successful. Cell morphology and growth rate were consistent from passage 10 onwards. Immunocytochemical examination of cellular proteins and matrix components confirmed the mechanical stability of the cells. Actin, fibronectin, vinculin, vimentin, and tubulin are present in the cells and form a network. In addition, the transport of molecules is ensured by the necessary proteins. Gene expression analysis showed a shift in the expressions of stem cell markers between younger and higher passages. While SOX2 and IGF1 were more highly expressed in the seventh passage, SOX9 and IGF2 expressions were significantly increased in higher passages. Therefore, the stable cell culture CMAfin1 can be used for applied analysis to further understand the cell physiology of C. maranea.

摘要

在本研究中,首次培育出了湖红点鲑(Coregonus maraena)的细胞系。湖红点鲑是一种濒危物种,研究表明,由于气候变化,该鱼类种群数量的进一步减少将对其产生影响。这个细胞系被命名为CMAfin1,在添加了10%胎牛血清的Leibovitz L - 15培养基中已维持培养超过3年。传代培养和保存(-80°C短期保存和液氮长期保存)均取得成功。从第10代起,细胞形态和生长速率保持一致。对细胞蛋白质和基质成分的免疫细胞化学检测证实了细胞的机械稳定性。细胞中存在肌动蛋白、纤连蛋白、纽蛋白、波形蛋白和微管蛋白,并形成一个网络。此外,必要的蛋白质确保了分子的运输。基因表达分析表明,干细胞标志物的表达在传代较年轻和传代较高的细胞之间存在差异。虽然SOX2和IGF1在第7代时表达较高,但SOX9和IGF2的表达在传代较高的细胞中显著增加。因此,稳定的细胞培养物CMAfin1可用于应用分析,以进一步了解湖红点鲑的细胞生理学。

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