| Live-cell Imaging of Neisseria meningitidis Microcolony Dispersal Induced by Lactate or Other Molecules
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Author:
Date:
2018-01-20
[Abstract] To efficiently colonize the nasopharyngeal epithelium, the human restricted pathogen Neisseria meningitidis follows a multistep adhesion cascade. First, the bacteria adhere to host cells and aggregate into spherical shaped structures called microcolonies. Several hours later, single bacteria start dispersing from the microcolonies and form a monolayer on top of the host cells. Once in proximity to host cells meningococci can adhere tightly to the epithelial surface or become internalized. This can eventually result in invasion of the mucosal surfaces and gain access to the bloodstream, causing a life-threatening disease. Lactate, a metabolite derived from human epithelial cells, has been previously shown to induce rapid dispersal of N. meningitidis from microcolonies. ...
[摘要] 为了有效地定居鼻咽上皮，人类限制性病原体脑膜炎奈瑟氏球菌遵循多步粘附级联。首先，细菌粘附到宿主细胞并聚集成称为微菌落的球形结构。几个小时后，单细菌开始从微菌落分散并在宿主细胞上形成单层。一旦接近宿主细胞，脑膜炎球菌可紧密地粘附在上皮表面或内化。这最终可能导致粘膜表面的侵入并进入血液，导致危及生命的疾病。乳酸是一种来源于人类上皮细胞的代谢物，之前已被证明能诱导N的快速分散。 meningitidis 来自微菌落。在这里，我们描述基于活细胞成像的宿主细胞自由方法来检查宿主来源的乳酸对N的时间的影响。 meningitides microcolony扩散。虽然在这个协议中我们使用乳酸盐，它可以很容易地修改，以测试其他分子的影响。
【背景】ñ。脑膜炎是引起败血症和/或脑膜炎的专性人类病原体。鼻咽上皮的初始附着和随后形成的小菌落是建立感染的第一步。为了导致疾病N。脑膜炎奈瑟球必须穿过鼻咽粘膜上皮屏障，其天然储库，并进入血液（Stephens，2009; ...
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| Generation of Tumour-stroma Minispheroids for Drug Efficacy Testing
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Author:
Date:
2017-01-05
[Abstract] The three-dimensional organisation of cells in a tissue and their interaction with adjacent cells and extracellular matrix is a key determinant of cellular responses, including how tumour cells respond to stress conditions or therapeutic drugs (Elliott and Yuan, 2011). In vivo, tumour cells are embedded in a stroma formed primarily by fibroblasts that produce an extracellular matrix and enwoven with blood vessels. The 3D mixed cell type spheroid model described here incorporates these key features of the tissue microenvironment that in vivo tumours exist in; namely the three-dimensional organisation, the most abundant stromal cell types (fibroblasts and endothelial cells), and extracellular matrix. This method combined with confocal microscopy can be a powerful tool to ...
[摘要] 组织中细胞的三维组织及其与相邻细胞和细胞外基质的相互作用是细胞反应的关键决定因素，包括肿瘤细胞如何对应激条件或治疗药物的反应（Elliott和Yuan，2011）。在体内，肿瘤细胞被包埋在主要由成纤维细胞形成的基质中，所述成纤维细胞产生细胞外基质并用血管编织。这里描述的3D混合细胞类型球体模型包括了体内存在肿瘤的组织微环境的这些关键特征;即三维组织，最丰富的基质细胞类型（成纤维细胞和内皮细胞）和细胞外基质。该方法结合共聚焦显微镜可以成为不同肿瘤类型的药物敏感性，血管生成和细胞迁移/侵袭测定的有力工具。
背景 传统的单层细胞培养（二维）强化人造环境，其与体内存在的组织细胞大不相同。最重要的区别之一是在单层培养物中，细胞是极化的，即，面向培养物的细胞表面和暴露于培养基的上细胞表面完全不同，经常反对的信号（Fitzgerald等人，2015）。为了解决细胞极化的问题，肿瘤球状体培养越来越多地用于癌症研究。肿瘤球体可以通过减少其通常发生在单层培养物中的生长培养基的扩散和稀释，通过细胞因子和趋化因子复制存在于组织中的三维细胞 - 细胞相互作用和一定程度的旁分泌信号传导（Lawlor等，2002; ...
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