Song Yuting, Shao Lijun, Yu Xiaoli
College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China.
Institute of Evolution & Marine Biodiversity, Ocean University of China, Qingdao 266003, China.
Biology (Basel). 2024 Jul 19;13(7):545. doi: 10.3390/biology13070545.
As an important research tool, cell lines play a vital role in life science research, medical research, and drug development. During the culture of the head kidney (TK) cell line, we found a phenomenon of cell vacuolization caused by excessive serum concentration. Moreover, the vacuolization of the cells gradually disappeared after passage by trypsin digestion. In clarifying the formation mechanism of this reversible cellular vacuolation, transcriptomics was utilized to explore the mechanism of cell vacuolization caused by excessive serum concentration. Transcriptome analysis indicated that excessive serum concentration could cause the up-regulated expression of PORCN and other genes to promote cell proliferation. Compared with cells whose vacuolization disappeared after trypsin digestion and passage, the expression of mitosis-related genes (BUB1, ttk, Mad2, Cdc20, CDK1, CCNB1), nuclear stability-related genes LMNB1 and tissue stress and repair-related genes HMMR in vacuolated cells caused by excessive serum concentration was significantly up-regulated. There is a regulatory system related to adaptation and stress repair in the cells, which can maintain cell stability to a certain extent. This study provides a theoretical basis for the stable culture of fish cell lines and the solution to the problem of cell vacuolation.
作为一种重要的研究工具，细胞系在生命科学研究、医学研究和药物开发中发挥着至关重要的作用。在头肾（TK）细胞系培养过程中，我们发现了因血清浓度过高导致细胞空泡化的现象。此外，经胰蛋白酶消化传代后，细胞的空泡化逐渐消失。在阐明这种可逆性细胞空泡化的形成机制时，利用转录组学来探究血清浓度过高导致细胞空泡化的机制。转录组分析表明，血清浓度过高会导致PORCN等基因表达上调以促进细胞增殖。与经胰蛋白酶消化传代后空泡化消失的细胞相比，血清浓度过高导致空泡化的细胞中，有丝分裂相关基因（BUB1、ttk、Mad2、Cdc20、CDK1、CCNB1）、核稳定性相关基因LMNB1以及组织应激与修复相关基因HMMR的表达显著上调。细胞内存在一个与适应和应激修复相关的调节系统，其可在一定程度上维持细胞稳定性。本研究为鱼类细胞系的稳定培养及细胞空泡化问题的解决提供了理论依据。
