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利用人输卵管器官培养(FTOC)和原代输卵管上皮细胞(FTEC)研究淋病奈瑟菌感染的生物学特性。

Use of Human Fallopian Tube Organ in Culture (FTOC) and Primary Fallopian Tube Epithelial Cells (FTEC) to Study the Biology of Neisseria gonorrhoeae Infection.

作者信息

Álamos-Musre A Said, Escobar Alejandro, Tapia Cecilia V, Christodoulides Myron, Rodas Paula I

机构信息

Laboratory of Medical Microbiology and Pathogenesis, Faculty of Medicine, Universidad Andres Bello, Concepción, Región del Bío-Bío, Chile.

Laboratorio Biología celular y molecular, Instituto de Ciencias Odontológicas, Facultad de Odontología, Universidad de Chile, Santiago, Región Metropolitana, Chile.

出版信息

Methods Mol Biol. 2019;1997:377-402. doi: 10.1007/978-1-4939-9496-0_22.

Abstract

Epithelial cells represent one of the most important physical barriers to many bacterial pathogens. In the case of Neisseria gonorrhoeae, the epithelial cell response is critical because they are the main target of the tissue damage triggered by the pathogen, particularly when the organism reaches the Fallopian tube (FT). Although the irreversible damage triggered by N. gonorrhoeae in the FT has been previously reported (ectopic pregnancy, pelvic inflammatory disease and infertility), the mechanisms of gonococcal-induced tissue damage are not fully understood. In addition, the lack of animal models that efficiently mimic the human disease and the complexity of gonococcus-host interactions make studying gonococcal pathogenesis particularly difficult. The use of human immortalized cells is also limited, since a variety of commercial FT cell lines is not yet available. Finally, the phase and antigenic variation of many gonococcal surface molecules involved in attachment and invasion of epithelial tissues leads to a failure to reproduce results using different human cells lines used in previous studies. The FT organ in culture (FTOC) and primary human fallopian tube epithelial cell (FTEC) represent the closest ex vivo cell models to explore the biology of Neisseria gonorrhoeae during infection of the FT, since it is a natural host target of the gonococcus. In this chapter, we describe protocols to process human FT samples to obtain FTOC and FTEC and assess their response to infection with Neisseria gonorrhoeae.

摘要

上皮细胞是许多细菌病原体最重要的物理屏障之一。就淋病奈瑟菌而言,上皮细胞反应至关重要,因为它们是该病原体引发组织损伤的主要靶点,尤其是当病原体到达输卵管(FT)时。尽管先前已报道淋病奈瑟菌在输卵管中引发的不可逆损伤(异位妊娠、盆腔炎和不孕症),但其诱导组织损伤的机制尚未完全了解。此外,缺乏能有效模拟人类疾病的动物模型以及淋病奈瑟菌与宿主相互作用的复杂性使得研究淋病奈瑟菌发病机制尤为困难。人类永生化细胞的使用也受到限制,因为目前尚无多种商业化的输卵管细胞系。最后,许多参与上皮组织附着和侵袭的淋病奈瑟菌表面分子的相位和抗原变异导致无法使用先前研究中使用的不同人类细胞系重现结果。培养中的输卵管器官(FTOC)和原代人输卵管上皮细胞(FTEC)是探索淋病奈瑟菌感染输卵管期间生物学特性最接近的体外细胞模型,因为输卵管是淋病奈瑟菌的天然宿主靶点。在本章中,我们描述了处理人类输卵管样本以获得FTOC和FTEC并评估它们对淋病奈瑟菌感染反应的方案。

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