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Comparison of Chlamydia trachomatis serovar L2 growth in polarized genital epithelial cells grown in three-dimensional culture with non-polarized cells

机译:沙眼衣原体血清型L2在三维培养的非生殖细胞极化生殖器上皮细胞中的生长比较

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摘要

A common model for studying Chlamydia trachomatis and growing chlamydial stocks uses Lymphogranuloma venereum serovar L2 and non-polarized HeLa cells. However, recent publications indicate that the growth rate and progeny yields can vary considerably for a particular strain depending on the cell line/type used, and seem to be partially related to cell tropism. In the present study, the growth of invasive serovar L2 was compared in endometrial HEC-1B and endocervical HeLa cells polarized on collagen-coated microcarrier beads, as well as in HeLa cells grown in tissue culture flasks. Microscopy analysis revealed no difference in chlamydial attachment/entry patterns or in inclusion development throughout the developmental cycle between cell lines. Very comparable growth curves in both cell lines were also found using real-time PCR analysis, with increases in chlamydial DNA content of 400–500-fold between 2 and 36 h post-inoculation. Similar progeny yields with comparable infectivity were recovered from HEC-1B and HeLa cell bead cultures, and no difference in chlamydial growth was found in polarized vs. non-polarized HeLa cells. In conclusion, unlike other C. trachomatis strains such as urogenital serovar E, invasive serovar L2 grows equally well in physiologically different endometrial and endocervical environments, regardless of the host cell polarization state.
机译:研究沙眼衣原体和生长衣原体种群的常用模型是使用性淋巴肉芽肿性病毒L2血清和非极化HeLa细胞。但是,最近的出版物表明,对于特定菌株,其生长速率和后代产量可能会有所不同,具体取决于所使用的细胞系/类型,并且似乎与细胞嗜性部分相关。在本研究中,比较了在涂有胶原蛋白的微载体珠粒上极化的子宫内膜HEC-1B和宫颈内膜HeLa细胞以及组织培养瓶中生长的HeLa细胞中侵袭性血清型L2的生长。显微镜分析表明,在整个细胞系之间的整个发育周期中,衣原体附着/进入模式或包涵体形成没有差异。实时PCR分析还发现两种细胞系中的生长曲线具有可比性,接种后2至36小时内衣原体DNA含量增加了400-500倍。从HEC-1B和HeLa细胞微珠培养物中回收到具有相似传染性的相似子代产量,并且在极化与非极化HeLa细胞中未发现衣原体生长的差异。总之,与其他沙眼衣原体菌株(如泌尿生殖系统血清型E)不同,无论宿主细胞的极化状态如何,侵入性血清型L2在生理上不同的子宫内膜和宫颈内环境中均能生长良好。

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