Chen Phylip, K C Mahesh, Peeples Mark E
Center for Microbe and Immunity Research, Abigail Wexner Research Institute, Nationwide Children's Hospital, Columbus, OH, USA.
Methods Mol Biol. 2025;2948:249-265. doi: 10.1007/978-1-0716-4666-3_16.
Human viruses have traditionally been studied in immortalized, usually tumor-derived, cell lines because of their availability, low cost, ease of expansion, and ease of care. Although most viruses infect, replicate, and spread in these cells, all aspects of in vivo virus-host interaction are not accurately reflected in immortalized cells. The isolation, storage, and differentiation of human airway epithelial basal cells have enabled ex vivo studies of RSV infection in a near natural setting. Here we provide our rationale for using well-differentiated primary human bronchial and nasal epithelial cultures, and a current protocol for generating these fully differentiated airway cultures from donor tissue for ex vivo studies of RSV and other respiratory virus infections.
传统上,人类病毒是在永生化细胞系中进行研究的,这些细胞系通常来源于肿瘤,因为它们容易获得、成本低、易于扩增且易于培养。尽管大多数病毒能在这些细胞中感染、复制和传播,但永生化细胞并不能准确反映体内病毒与宿主相互作用的所有方面。人呼吸道上皮基底细胞的分离、储存和分化,使得在接近自然的环境中对呼吸道合胞病毒(RSV)感染进行离体研究成为可能。在此,我们阐述了使用高度分化的原代人支气管和鼻上皮培养物的理论依据,并提供了一个当前的方案,用于从供体组织生成这些完全分化的气道培养物,以用于RSV和其他呼吸道病毒感染的离体研究。