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首页> 外文期刊>Cilia >Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium: a simple methodological approach for ex vivo studies of primary ciliary dyskinesia
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Patient-specific three-dimensional explant spheroids derived from human nasal airway epithelium: a simple methodological approach for ex vivo studies of primary ciliary dyskinesia

机译：源自人鼻气道上皮的患者特定三维外植体球体：一种简单的方法学方法，用于原发性睫状运动障碍的离体研究

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Background Three-dimensional explant spheroid formation is an ex vivo technique previously used in studies of airway epithelial ion and water transport. Explanted cells and sheets of nasal epithelium form fully differentiated spheroids enclosing a partly fluid-filled lumen with the ciliated apical surface facing the outside and accessible for analysis of ciliary function. Methods We performed a two-group comparison study of ciliary beat pattern and ciliary beat frequency in spheroids derived from nasal airway epithelium in patients with primary ciliary dyskinesia (PCD) and in healthy controls. Nasal ciliary cells and sheets were removed on day 1 by nasal brush biopsy and analyzed with regard to ciliary beat pattern—and frequency using high-speed video imaging for standard reference values. Three-dimensional explant spheroid formation was initiated in the same individual on the same day by incubation of cells and sheets from a separate brush biopsy. Harvested spheroids were analyzed earliest possible and values of spheroid ciliary beat pattern and frequency were compared to the corresponding reference values from day 1. Results Spheroids formed fast in serum-free culture medium. Formation was successful in 15 out of 18 (82%) sampled individuals. Thus, formation was successful in seven healthy controls and eight PCD patients, while unsuccessful in 3 with PCD due to infection. Median (range) number of days in culture before harvesting of spheroids was 4 (1–5) in healthy versus 2 (1–5) in PCD. Spheroid ciliary beat pattern and frequency were unchanged compared to their corresponding day 1 standard reference values. Spheroid ciliary beat frequency discriminated highly significant between healthy controls (9.3?Hz) and PCD patients (2.4?Hz) ( P Conclusion Patient-specific three-dimensional explant spheroid formation from a minimal invasive nasal brush biopsy is a feasible, fast and valid ex vivo method to assess ciliary function with potential of aiding the diagnosis of PCD. In addition, it may be a useful model in the investigation of pathophysiological aspects and drug effects in human nasal airway epithelium.
机译：背景技术三维外植体球体形成是一种先前用于气道上皮离子和水传输研究的离体技术。移出的细胞和鼻上皮细胞片形成完全分化的球状体，包围部分充满液体的管腔，纤毛的顶表面朝外，可用于分析纤毛功能。方法我们对原发性睫状运动障碍（PCD）患者和健康对照者的鼻气道上皮球状体的纤毛跳动模式和纤毛跳动频率进行了两组比较研究。在第1天，通过鼻刷活检取出鼻睫状细胞和片状细胞，并使用高速视频成像作为标准参考值，分析睫状跳动模式和频率。通过孵育来自单独刷式活检的细胞和薄片，在同一个人的同一天开始三维外植体球体的形成。尽早分析收获的球状体，并将球状体纤毛节律模式和频率值与第1天开始的相应参考值进行比较。结果球状体在无血清培养基中快速形成。 18个样本中有15个样本（82％）成功形成。因此，在7名健康对照和8名PCD患者中成功形成，而在3名因感染PCD中未成功。健康人收获球体之前的培养天数（范围）为4（1-5），而PCD为2（1-5）。球状纤毛的搏动模式和频率与其相应的第1天标准参考值相比没有变化。在健康对照组（9.3？Hz）和PCD患者（2.4？Hz）之间，球状睫状心跳频率有很高的区别（P结论通过微创鼻刷活检对患者进行特定的三维外植体球状形成是可行，快速且有效的评估睫状功能的体内方法可能有助于PCD的诊断，此外，它可能是研究人鼻气道上皮的病理生理方面和药物作用的有用模型。

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