Cell line development from shrimp is not a novel venture as researchers across the globe have been trying to have crustacean cell lines over 30 years. The reason for not attaining a crustacean or precisely a shrimp cell line is believed to be the replicative senescence and the inability to maintain telomere length in vitro. Moreover, spontaneous in vitro transformations do not happen in shrimp cells. Oncogenic induction in primary cell culture is one of the ways to attain in vitro transformation by way of disrupting the mechanisms which involve cellular senescence. In this context, a recombinant baculovirus with shrimp viral promoter IHHNV-P2 was used for the transduction aimed at immortalization. An oncogene, H-ras, was successfully amplified and cloned in to the baculoviral vector, downstream to shrimp viral promoter IHHNV-P2 and upstream to GFP. Recombinant baculovirus with H-ras was generated and used for transduction into shrimp lymphoid cells during early dividing stage. Accordingly, fibroblast-like primary cell culture got developed, and H-ras and GFP expression could be confirmed. The study suggests that the simple method of incubating recombinant baculovirus with minced tissue enables in vitro transduction during early dividing stage of the cells, and the transduction efficiency gets enhanced by adding 5 mM sodium butyrate to the culture medium.

从虾中开发细胞系并不是一项新的冒险，因为 30 多年来，全球研究人员一直在尝试开发甲壳类动物细胞系。没有获得甲壳类动物或确切地说是虾细胞系的原因被认为是复制衰老和无法在体外维持端粒长度。此外，在虾细胞中不会发生自发的体外转化。原代细胞培养中的致癌诱导是通过破坏涉及细胞衰老的机制来实现体外转化的方法之一。在这种情况下，具有虾病毒启动子 IHHNV-P2 的重组杆状病毒用于旨在永生化的转导。致癌基因 H- ras, 被成功扩增并克隆到杆状病毒载体中，下游到虾病毒启动子 IHHNV-P2 和上游到 GFP。产生了带有 H- ras 的重组杆状病毒，并用于在早期分裂阶段转导入虾淋巴细胞。因此，成纤维细胞样原代细胞培养物得到发展，并且可以确认H- ras和GFP表达。该研究表明，重组杆状病毒与切碎的组织一起孵育的简单方法可以在细胞分裂的早期进行体外转导，并且通过向培养基中加入 5 mM 丁酸钠可以提高转导效率。