[Establishment of human sapovirus culture method].

Uirusu Pub Date : 2023-01-01 DOI:10.2222/jsv.73.1
Hirotaka Takagi, Tomoichiro Oka
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Abstract

More than 40 years after the discovery of human sapovirus (HuSaV), we have established a HuSaV culture system in which HuTu80 cells derived from the human duodenum adenocarcinoma cell line are cultured together with the addition of bile acid as a supplement. In addition to being a common cell line, this system using HuTu80 cells is a versatile method because classical culture media are available, and it is easy to scale-up for culture. However, the number of culture days required to obtain sufficient viral titer, the confirmation of viral gene conservation for sample selection, and the method for passaging of HuTu80-cells were crucial. So far, 15 genotypes have been successfully propagated and stocked, and stable supply as research resources has been achieved. Due to the above efforts, we can now proceed with the production and analysis of antisera using purified antigens and the evaluation of inactivation conditions. This manuscript introduces the background for selection of the cell line and bile acids, and the topics that have been discussed since the publication, as well as future issues that were raised such as the expression of cytopathicity and elucidation of low UV-C sensitivity of fecal-derived HuSaV.

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[建立人类沙波病毒培养方法]。
在人类沙波病毒(HuSaV)被发现 40 多年后,我们建立了一种 HuSaV 培养系统,在该系统中,从人类十二指肠腺癌细胞系中提取的 HuTu80 细胞与添加胆汁酸作为辅料的细胞一起培养。除了是一种常见的细胞系外,这种使用 HuTu80 细胞的系统还是一种通用的方法,因为可以获得经典的培养基,而且易于扩大培养规模。然而,获得足够病毒滴度所需的培养天数、用于样本选择的病毒基因保护的确认以及 HuTu80 细胞的传代方法都至关重要。到目前为止,已成功繁殖和储备了 15 个基因型,并实现了作为研究资源的稳定供应。由于上述努力,我们现在可以使用纯化的抗原生产和分析抗血清,并对灭活条件进行评估。本手稿介绍了细胞系和胆汁酸的选择背景、发表后的讨论主题以及今后提出的问题,如细胞病理学的表达和粪源性 HuSaV 对紫外线-C 低敏感性的阐明。
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