Abstract：
The emergence of the coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) inspired rapid research efforts targeting the host range, pathogenesis and transmission mechanisms, and the development of antiviral strategies. Genetically modified mice, rhesus macaques, ferrets, and Syrian golden hamsters have been frequently used in studies of pathogenesis and efficacy of antiviral compounds and vaccines. However, alternatives to in vivo experiments, such as immortalized cell lines, primary respiratory epithelial cells cultured at an air-liquid interface, stem/progenitor cell-derived organoids, or tissue explants, have also been used for isolation of SARS-CoV-2, investigation of cytopathic effects, and pathogen-host interactions. Moreover, initial proof-of-concept studies for testing therapeutic agents can be performed with these tools, showing that animal-sparing cell culture methods could significantly reduce the need for animal models in the future, following the 3R principles of replace, reduce, and refine. So far, only few studies using animal-derived primary cells or tissues have been conducted in SARS-CoV-2 research, although natural infection has been shown to occur in several animal species. Therefore, the need for in-depth investigations on possible interspecies transmission routes and differences in susceptibility to SARS-CoV-2 is urgent. This review gives an overview of studies employing alternative culture systems like primary cell cultures, tissue explants, or organoids for investigations of the pathophysiology and reverse zoonotic potential of SARS-CoV-2 in animals. In addition, future possibilities of SARS-CoV-2 research in animals, including previously neglected methods like the use of precision-cut lung slices, will be outlined.
摘要：
由严重急性呼吸道综合症冠状病毒2（SARS-CoV-2）引起的2019年冠状病毒病（COVID-19）的出现激发了针对宿主范围的快速研究工作，发病机制和传播机制，以及抗病毒策略的发展。转基因小鼠,猕猴,雪貂,和叙利亚金仓鼠经常用于抗病毒化合物和疫苗的发病机理和功效的研究。然而,替代体内实验，如永生化细胞系，在气-液界面培养的原代呼吸道上皮细胞，干细胞/祖细胞衍生的类器官，或组织外植体，还用于分离SARS-CoV-2，研究细胞病变效应，和病原体-宿主相互作用。此外，可以使用这些工具进行测试治疗剂的初步概念验证研究，这表明节省动物的细胞培养方法可以显着减少未来对动物模型的需求，遵循3R替换原则，reduce,和精炼。到目前为止,在SARS-CoV-2研究中，使用动物来源的原代细胞或组织进行的研究很少，尽管已经证明自然感染发生在几种动物物种中。因此,迫切需要对可能的种间传播途径和SARS-CoV-2易感性差异进行深入研究。这篇综述概述了采用替代培养系统如原代细胞培养的研究，组织外植体，或类器官，用于研究动物中SARS-CoV-2的病理生理学和反向人畜共患病潜力。此外,SARS-CoV-2在动物中研究的未来可能性，包括以前被忽视的方法，如使用精确切割的肺切片，将被概述。
