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蝙蝠气道上皮细胞:一种用于研究人畜共患病毒的新型工具。

Bat airway epithelial cells: a novel tool for the study of zoonotic viruses.

作者信息

Eckerle Isabella, Ehlen Lukas, Kallies René, Wollny Robert, Corman Victor M, Cottontail Veronika M, Tschapka Marco, Oppong Samuel, Drosten Christian, Müller Marcel A

机构信息

Institute of Virology, University of Bonn Medical Centre, Bonn, Germany.

Institute of Experimental Ecology, University of Ulm, Ulm, Germany.

出版信息

PLoS One. 2014 Jan 13;9(1):e84679. doi: 10.1371/journal.pone.0084679. eCollection 2014.

Abstract

Bats have been increasingly recognized as reservoir of important zoonotic viruses. However, until now many attempts to isolate bat-borne viruses in cell culture have been unsuccessful. Further, experimental studies on reservoir host species have been limited by the difficulty of rearing these species. The epithelium of the respiratory tract plays a central role during airborne transmission, as it is the first tissue encountered by viral particles. Although several cell lines from bats were established recently, no well-characterized, selectively cultured airway epithelial cells were available so far. Here, primary cells and immortalized cell lines from bats of the two important suborders Yangochiroptera and Yinpterochiroptera, Carollia perspicillata (Seba's short-tailed bat) and Eidolon helvum (Straw-colored fruit bat), were successfully cultured under standardized conditions from both fresh and frozen organ specimens by cell outgrowth of organ explants and by the use of serum-free primary cell culture medium. Cells were immortalized to generate permanent cell lines. Cells were characterized for their epithelial properties such as expression of cytokeratin and tight junctions proteins and permissiveness for viral infection with Rift-Valley fever virus and vesicular stomatitis virus Indiana. These cells can serve as suitable models for the study of bat-borne viruses and complement cell culture models for virus infection in human airway epithelial cells.

摘要

蝙蝠越来越被认为是重要人畜共患病毒的宿主。然而,到目前为止,许多在细胞培养中分离蝙蝠携带病毒的尝试都未成功。此外,对宿主物种的实验研究受到饲养这些物种困难的限制。呼吸道上皮在空气传播过程中起核心作用,因为它是病毒颗粒遇到的第一个组织。尽管最近建立了几种蝙蝠细胞系,但到目前为止还没有特征明确、可选择性培养的气道上皮细胞。在这里,通过器官外植体的细胞生长和使用无血清原代细胞培养基,成功地在标准化条件下从新鲜和冷冻的器官标本中培养了来自两个重要亚目——阳翼手亚目和阴翼手亚目的蝙蝠的原代细胞和永生化细胞系,即秘鲁叶鼻蝠(塞氏短尾蝠)和锤头果蝠。细胞被永生化以产生永久细胞系。对细胞的上皮特性进行了表征,如细胞角蛋白和紧密连接蛋白的表达,以及对裂谷热病毒和印第安纳水疱性口炎病毒感染的易感性。这些细胞可作为研究蝙蝠携带病毒的合适模型,并补充人类气道上皮细胞病毒感染的细胞培养模型。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41c0/3890267/de3b2d744ee6/pone.0084679.g001.jpg

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