Immunocompetent 3D model of human upper airway for disease modeling and in vitro drug evaluation.
Immunocompetent 3D model of human upper airway for disease modeling and in vitro drug evaluation.
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DOI：
10.1021/mp5000295
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发表时间：
2014-07-07
影响因子：
4.9
通讯作者：
Ghaemmaghami AM
中科院分区：
文献类型：
--
作者：
Harrington H;Cato P;Salazar F;Wilkinson M;Knox A;Haycock JW;Rose F;Aylott JW;Ghaemmaghami AM
由于动物模型与人类的生物学相关性有限以及伦理方面的考虑，需要开发更复杂的体外模型来评估新药和化学品。尽管存在一些基于人体细胞的分析，但它们通常是2D的，由单一细胞类型组成，并且对天然组织的细胞和功能代表有限。在这项研究中，我们使用仿生多孔电纺支架来开发一个具有免疫活性的人体呼吸道3D模型，该模型由存在于上呼吸道上皮中的三种关键细胞类型组成。这三种细胞类型，即上皮细胞(提供物理屏障)、成纤维细胞(产生细胞外基质)和树突状细胞(免疫传感)，最初生长在单独的支架上，然后组装成3D多细胞组织模型。上皮层在气液界面培养长达四周，形成功能性屏障，表现为跨上皮电阻(TEER)增加和紧密连接的形成。通过量化TEER读数的变化和通过免疫染色评估细胞紧密连接来监测上皮细胞对过敏原暴露的反应。研究发现，与成纤维细胞共培养的上皮细胞形成功能性上皮屏障的速度比单独培养上皮细胞的速度更快，从过敏原暴露中恢复也更快。此外，我们的数据显示，这个模型中的树突状细胞仍然活着，并对外部刺激做出反应，这一点从它们在3D结构中对过敏原挑战的反应中迁移得到了证明。该模型提供了一个易于组装和生理相关的人体呼吸道上皮3D模型，可用于更好地了解肺部生物学、免疫细胞之间的相互作用、空气传播的过敏原和病原体以及药物输送的研究。
The development of more complex in vitro models for the assessment of novel drugs and chemicals is needed because of the limited biological relevance of animal models to humans as well as ethical considerations. Although some human-cell-based assays exist, they are usually 2D, consist of single cell type, and have limited cellular and functional representation of the native tissue. In this study, we have used biomimetic porous electrospun scaffolds to develop an immunocompetent 3D model of the human respiratory tract comprised of three key cell types present in upper airway epithelium. The three cell types, namely, epithelial cells (providing a physical barrier), fibroblasts (extracellular matrix production), and dendritic cells (immune sensing), were initially grown on individual scaffolds and then assembled into the 3D multicell tissue model. The epithelial layer was cultured at the air–liquid interface for up to four weeks, leading to formation of a functional barrier as evidenced by an increase in transepithelial electrical resistance (TEER) and tight junction formation. The response of epithelial cells to allergen exposure was monitored by quantifying changes in TEER readings and by assessment of cellular tight junctions using immunostaining. It was found that epithelial cells cocultured with fibroblasts formed a functional epithelial barrier at a quicker rate than single cultures of epithelial cells and that the recovery from allergen exposure was also more rapid. Also, our data show that dendritic cells within this model remain viable and responsive to external stimulation as evidenced by their migration within the 3D construct in response to allergen challenge. This model provides an easy to assemble and physiologically relevant 3D model of human airway epithelium that can be used for studies aiming at better understanding lung biology, the cross-talk between immune cells, and airborne allergens and pathogens as well as drug delivery.
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影响因子：
14
作者：
Ma, ZW;Kotaki, M;Ramakrishna, S
通讯作者：
Ramakrishna, S
影响因子：
7.4
作者：
Ghaemmaghami, Amir M.;Hancock, Matthew J.;Harrington, Helen;Kaji, Hirokazu;Khademhosseini, Ali
通讯作者：
Khademhosseini, Ali
影响因子：
14
作者：
Bashur, Chris A.;Dahlgren, Linda A.;Goldstein, Aaron S.
通讯作者：
Goldstein, Aaron S.
影响因子：
9.6
作者：
Bergeron, C;Boulet, LP
通讯作者：
Boulet, LP
影响因子：
9
作者：
Chau, David Y. S.;Johnson, Claire;Ghaemmaghami, Amir M.
通讯作者：
Ghaemmaghami, Amir M.