Abstract Continuous shrimp cell cultures offer opportunities for studying shrimp pathogens at the cellular and molecular level and developing diagnostic tools. However, no continuous shrimp cell lines have yet been successfully established. This might be due to the lack of information on the molecular mechanisms that control shrimp cell proliferation and cell arrest under culture conditions. In this study, differentially expressed genes (DEGs) in shrimp primary culture cells were determined by comparison with normal tissue (in vivo). This study aimed to comprehensively identify key regulator genes that control cell proliferation and cell cycle arrest in culture conditions. The primary shrimp cells were derived from testicular tissue and cultured in vitro. RNA sequencing (RNA-Seq) was performed to investigate change in gene expression level across the entire transcriptome between shrimp primary cells and normal tissue (in vivo). RNA-seq results revealed over 100 genes with distinct gene expression patterns between primary cells and normal tissue. The DEG results and functional gene analysis showed a clear difference in gene expression patterns of cell cycle-related genes between primary cells and normal tissue. LRWD1, TMEM127, CDCA3, PPP2R1A, and GOLGA2, which are cell cycle-related genes, were downregulated in shrimp primary cells. These genes are required for G1/S phase transition, entry into mitosis, and cell proliferation control. However, a gene that induces cell cycle arrest, ARAF, was upregulated in culture conditions. The results demonstrated that these genes might play essential roles in cell proliferation and cell arrest under culture conditions. These candidate genes may be alternative potential targets for genetic manipulation to maintain cell proliferation and establishment of a shrimp continuous cell line.

中文翻译：

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基于体外培养对虾原代细胞基因表达的影响，研究必需细胞周期调节基因作为永生化虾细胞系建立的候选基因

摘要 连续虾细胞培养为在细胞和分子水平上研究虾病原体和开发诊断工具提供了机会。然而，尚未成功建立连续的虾细胞系。这可能是由于缺乏关于在养殖条件下控制虾细胞增殖和细胞停滞的分子机制的信息。在这项研究中，通过与正常组织（体内）比较来确定虾原代培养细胞中的差异表达基因（DEG）。本研究旨在全面鉴定在培养条件下控制细胞增殖和细胞周期停滞的关键调节基因。原代虾细胞来源于睾丸组织并在体外培养。进行 RNA 测序 (RNA-Seq) 以研究虾原代细胞和正常组织（体内）之间整个转录组中基因表达水平的变化。RNA-seq 结果揭示了 100 多个基因，它们在原代细胞和正常组织之间具有不同的基因表达模式。DEG结果和功能基因分析显示原代细胞与正常组织细胞周期相关基因的基因表达模式存在明显差异。LRWD1、TMEM127、CDCA3、PPP2R1A 和 GOLGA2 是细胞周期相关基因，在虾原代细胞中下调。这些基因是 G1/S 期转变、进入有丝分裂和细胞增殖控制所必需的。然而，诱导细胞周期停滞的基因 ARAF 在培养条件下被上调。结果表明，这些基因可能在培养条件下的细胞增殖和细胞停滞中发挥重要作用。这些候选基因可能是基因操作的替代潜在目标，以维持细胞增殖和建立虾连续细胞系。