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首页> 外文期刊>Journal of Epithelial Biology & Pharmacology >Micropatterning Co-cultures of Epithelial Cells on Filter Insert Substrates
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Micropatterning Co-cultures of Epithelial Cells on Filter Insert Substrates

机译：滤器插入基质上皮细胞的微模式共培养

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A key feature of mature epithelium is the presence of an apical-basal polarization. In vitro this is achieved byculturing either pure populations of epithelial cells or unorganized mixtures of primary cell populations harvested fromepithelial tissues in a filter insert culture system. In addition to epithelial cells however, multiple other cell types are alsopresent in normal epithelium in a specific organization. Generally the organization of component cells within a tissue iscritical for normal function. The importance of the organization of different cell populations within normal and diseasedepithelium in vitro has received little attention, in part due to the lack of available methods for spatially controllingmultiple cell populations while maintaining the culture conditions necessary to generate polarized and mature epithelium.We have developed a set of micropatterning techniques to spatially control the organization of co-cultures of epithelialcells on filter insert membranes under the culture conditions necessary to induce epithelial cell polarization. We describe afibronectin microprinting method that allows arbitrary pattern generation and a parafilm insert method that allows onlysimple stripe geometry patterning but does not require the photolithographic equipment that the fibronectin microprintingmethod requires, and can be adopted by any epithelial biology lab. We also demonstrate that our micropatterning methodsdo not alter the formation of cilia, a marker of epithelium maturation. Our methods provide a novel tool for studyingepithelial biology in polarized epithelial sheets containing multiple cell populations with controlled organization.
机译：成熟的上皮细胞的一个关键特征是根尖极化。在体外，这可以通过在滤膜插入培养系统中培养上皮细胞的纯群体或从上皮组织收获的原代细胞群体的无组织混合物来实现。然而，除了上皮细胞以外，在特定组织中正常上皮中还存在多种其他细胞类型。通常，组织内组成细胞的组织对于正常功能至关重要。正常和疾病上皮细胞中不同细胞群体的组织重要性的关注很少受到关注，部分原因是缺乏在空间上控制多个细胞群体同时保持产生极化和成熟上皮细胞所需的培养条件的可用方法。一套微模式技术，用于在诱导上皮细胞极化所必需的培养条件下，空间控制滤器插入膜上的上皮细胞共培养的组织。我们描述了可以进行任意图案生成的纤连蛋白微打印方法和仅允许简单的条纹几何图案化但不需要纤连蛋白微打印方法所需的光刻设备的石蜡膜插入方法，并且可以被任何上皮生物学实验室采用。我们还证明了我们的微图案化方法不会改变纤毛的形成，纤毛是上皮成熟的标志。我们的方法提供了一种新的工具，用于研究极化的上皮细胞片中的上皮生物学，其中包含多个具有受控组织的细胞群。

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