A Synthetic Hydrogel, VitroGel(®) ORGANOID-3, Improves Immune Cell-Epithelial Interactions in a Tissue Chip Co-Culture Model of Human Gastric Organoids and Dendritic Cells.
A Synthetic Hydrogel, VitroGel(®) ORGANOID-3, Improves Immune Cell-Epithelial Interactions in a Tissue Chip Co-Culture Model of Human Gastric Organoids and Dendritic Cells.
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DOI：
10.3389/fphar.2021.707891
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发表时间：
2021
影响因子：
5.6
通讯作者：
Bimczok D
中科院分区：
文献类型：
--
作者：
Cherne MD;Sidar B;Sebrell TA;Sanchez HS;Heaton K;Kassama FJ;Roe MM;Gentry AB;Chang CB;Walk ST;Jutila M;Wilking JN;Bimczok D
单核吞噬细胞 (MNP) 对胃肠道上皮的免疫监视对于维持肠道健康至关重要。然而，研究人类胃肠道上皮和树突状细胞（DC）等 MNP 之间复杂的相互作用很困难，因为传统的细胞培养系统缺乏复杂性，并且动物模型可能无法充分代表人体组织。微生理系统或组织芯片是这些研究的一个有吸引力的替代方案，因为它们使用重建生理组织微环境的微型培养平台来模拟特定组织或器官的功能特征。然而，在组织芯片平台上成功整合多种组织类型以重现生理细胞间相互作用仍然是一个挑战。我们之前开发了一种组织芯片系统，即肠道类器官流式芯片 (GOFlowChip)，用于长期培养 3D 多能干细胞衍生的人类肠道类器官。在这里，我们优化了 GOFlowChip 平台，构建了复杂的微生理免疫细胞-上皮细胞共培养模型，以研究人胃中 DC-上皮相互作用。我们首先测试了不同的管道材料和芯片配置，以优化 GOFlowChip 上的 DC 加载，并证明在 GOFlowChip 上培养 DC 长达 20 小时不会影响 DC 激活状态或活力。然而，Transwell 趋化性测定和实时共聚焦成像显示，Matrigel（常用于类器官培养的细胞外基质 (ECM) 材料）可阻止 DC 向类器官迁移以及直接 MNP-上皮接触的建立。因此，我们接下来通过替代 ECM 材料（包括基质胶-胶原混合物和合成水凝胶）评估 DC 趋化性。基于多糖的合成水凝胶 VitroGel®-ORGANOID-3 (V-ORG-3) 能够显着增加 DC 通过基质的趋化性，支持类器官的存活和生长，并且不会显着改变 DC 的激活或活力。在 GOFlowChip 上，流入芯片的 DC 快速迁移通过 V-ORG 基质，到达嵌入芯片深处的类器官，DC 与胃类器官之间的相互作用增强。 DC 和 V-ORG-3 嵌入胃类器官成功集成到 GOFlowChip 平台中，现在可以对 MNP-上皮相互作用进行实时成像，并可以对胃肠道 MNP 和上皮细胞之间复杂的相互作用进行其他研究，以响应病原体、候选药物和粘膜疫苗。
Immunosurveillance of the gastrointestinal epithelium by mononuclear phagocytes (MNPs) is essential for maintaining gut health. However, studying the complex interplay between the human gastrointestinal epithelium and MNPs such as dendritic cells (DCs) is difficult, since traditional cell culture systems lack complexity, and animal models may not adequately represent human tissues. Microphysiological systems, or tissue chips, are an attractive alternative for these investigations, because they model functional features of specific tissues or organs using microscale culture platforms that recreate physiological tissue microenvironments. However, successful integration of multiple of tissue types on a tissue chip platform to reproduce physiological cell-cell interactions remains a challenge. We previously developed a tissue chip system, the gut organoid flow chip (GOFlowChip), for long term culture of 3-D pluripotent stem cell-derived human intestinal organoids. Here, we optimized the GOFlowChip platform to build a complex microphysiological immune-cell-epithelial cell co-culture model in order to study DC-epithelial interactions in human stomach. We first tested different tubing materials and chip configurations to optimize DC loading onto the GOFlowChip and demonstrated that DC culture on the GOFlowChip for up to 20 h did not impact DC activation status or viability. However, Transwell chemotaxis assays and live confocal imaging revealed that Matrigel, the extracellular matrix (ECM) material commonly used for organoid culture, prevented DC migration towards the organoids and the establishment of direct MNP-epithelial contacts. Therefore, we next evaluated DC chemotaxis through alternative ECM materials including Matrigel-collagen mixtures and synthetic hydrogels. A polysaccharide-based synthetic hydrogel, VitroGel®-ORGANOID-3 (V-ORG-3), enabled significantly increased DC chemotaxis through the matrix, supported organoid survival and growth, and did not significantly alter DC activation or viability. On the GOFlowChip, DCs that were flowed into the chip migrated rapidly through the V-ORG matrix and reached organoids embedded deep within the chip, with increased interactions between DCs and gastric organoids. The successful integration of DCs and V-ORG-3 embedded gastric organoids into the GOFlowChip platform now permits real-time imaging of MNP-epithelial interactions and other investigations of the complex interplay between gastrointestinal MNPs and epithelial cells in their response to pathogens, candidate drugs and mucosal vaccines.
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影响因子：
7.3
作者：
Cougoule C;Lastrucci C;Guiet R;Mascarau R;Meunier E;Lugo-Villarino G;Neyrolles O;Poincloux R;Maridonneau-Parini I
通讯作者：
Maridonneau-Parini I
影响因子：
3.7
作者：
Anguiano M;Castilla C;Maška M;Ederra C;Peláez R;Morales X;Muñoz-Arrieta G;Mujika M;Kozubek M;Muñoz-Barrutia A;Rouzaut A;Arana S;Garcia-Aznar JM;Ortiz-de-Solorzano C
通讯作者：
Ortiz-de-Solorzano C
影响因子：
--
作者：
Chakrabarti, Jayati;Holokai, Loryn;Zavros, Yana
通讯作者：
Zavros, Yana
影响因子：
29.4
作者：
Bimczok, Diane;Grams, Jayleen M.;Smith, Phillip D.
通讯作者：
Smith, Phillip D.
影响因子：
1.8
作者：
Federer C;Yoo M;Tan AC
通讯作者：
Tan AC