A comparison of three Peyer's patch "M-like" cell culture models: particle uptake, bacterial interaction, and epithelial histology
A comparison of three Peyer's patch "M-like" cell culture models: particle uptake, bacterial interaction, and epithelial histology
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DOI：
10.1016/j.ejpb.2017.07.013
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发表时间：
2017-10-01
影响因子：
4.9
通讯作者：
Brayden, David J.
中科院分区：
文献类型：
--
作者：
Ahmad, Tauseef;Gogarty, Martina;Brayden, David J.
肠道Peyer's patch (PP) microfold (M)细胞通过滤泡相关上皮（FAE）运输微生物和颗粒，作为粘膜免疫监视系统的一部分。在体外，人类m样细胞共培养模型被用作研究纳米颗粒中抗原摄取的筛选，但这些模型是劳动密集型的，并且存在实验室间的差异。我们比较了三种最成熟的过滤培养Caco-2/Raji B细胞共培养体系。这些是模型A (Kerneis et al., 1997)，模型B （Gullberg et al., 2000）和模型C （Des Rieux et al., 2007）。采用经上皮耐药（TEER）、[C-14]-甘露醇的表观通透系数（P-app）、M细胞样组织学、乳胶颗粒和鼠伤寒沙门菌易位等指标作为评价标准。每种共培养模型都显示出粒子易位的显著增加。与单培养相比，截短的微绒毛是它们最一致的特征。des Rieux等人（2007）开发的反向模型显示，与其他两种模型相比，TEER减少，P-app增加，同时粒子易位增加最多。Gullberg等人（2000）开发的正常导向模型是唯一一个持续显示鼠伤寒沙门氏菌易位增加的模型。通过在过滤器上涂上双层基质（TM）涂层，改变Raji B细胞的培养基喂养方式，并限制B细胞的传代次数，实现了对Gullberg模型B的改进，这反映在颗粒易位增加和组织学改善上。总之，这是第一次在一项研究中对所有三种设计进行比较，每种设计都显示了m样细胞的表型特征。模型C是最稳健的共培养，而模型B协议可以通过优化几个变量来改进，并且比两个反向模型建立起来更简单。(C) 2017 Elsevier B.V.版权所有
Intestinal Peyer's patch (PP) microfold (M) cells transport microbes and particulates across the follicle associated epithelium (FAE) as part of the mucosal immune surveillance system. In vitro human M-like cell co-culture models are used as screens to investigate uptake of antigens-in-nanoparticles, but the models are labour-intensive and there is inter-laboratory variability. We compared the three most established filter-grown Caco-2/Raji B cell co-culture systems. These were Model A (Kerneis et al., 1997), Model B (Gullberg et al., 2000), and Model C (Des Rieux et al. 2007). The criteria used were transepithelial resistance (TEER), the apparent permeability coefficient (P-app) of [C-14]-mannitol, M cell-like histology, as well as latex particle and Salmonella typhimurium translocation. Each co-culture model displayed substantial increases in particle translocation. Truncated microvilli compared to mono-cultures was their most consistent feature. The inverted model developed by des Rieux et al. (2007) displayed reductions in TEER and an increased (P-app), accompanied by the largest increase in particle translocation compared to the other two models. The normally-oriented model developed by Gullberg et al. (2000) was the only one to consistently display an increased translocation of Salmonella typhimurium. By applying a double Matrigelml (TM) coating on filters, altering the medium feeding regime for Raji B cells, and restricting the passage number of B cells, improvements to the Gullberg model B were achieved, as reflected by increased particle translocation and improved histology. In conclusion, this is the first time all three designs have been compared in one study and each displays phenotypic features of M-like cells. While Model C was the most robust co-culture, the Model B protocol could be improved by optimizing several variables and is less complicated to establish than the two inverted models. (C) 2017 Elsevier B.V. All rights reserved.