A Next Generation Outer Retinal Microphysiological System

摘要 诱导多能干细胞（iPSC）的发现为科学家提供了开发 从患有遗传性疾病的患者中分化组织，并使用这些体外模型来研究疾病 机制和测试药物和基因为基础的治疗。然而，目前的分化方案往往失败， 以准确再现天然组织中存在的细胞组织。例如，即使是国家的- 产生三维视网膜类器官的现有技术视网膜分化方案并不准确地 再现了存在于外层视网膜中的细胞间的相互作用。具体来说，脉络膜内皮细胞 细胞很少存在，并且从不在RPE下形成有组织的层。同样，与其形成一个 与新发育的光感受器并列的单层，RPE细胞通常被发现成簇，位于 神经视网膜的边缘在本申请中，我们描述了一个专注于开发的程序， 验证微生理学系统的设计，以准确地重建复杂的结构， 灌注的外层视网膜。具体来说，我们建议将联合收割机超高分辨率、最先进的3D打印技术 视网膜支架，iPSC技术和微流体技术，以开发下一代微生理系统 它概括了外层视网膜单位。完成本建议中概述的目标将有助于铺平道路 未来的研究重点是评估复杂的视网膜退行性疾病的病理生理学， 年龄相关性黄斑变性和开发新的药物和基因为基础的治疗。

ABSTRACT The discovery of induced pluripotent stem cells (iPSCs) provided scientists with the opportunity to develop differentiated tissues from patients with inherited disease, and to use these in vitro models to study disease mechanisms and to test drug and gene-based therapeutics. However, current differentiation protocols often fail to accurately recapitulate the cellular organization present in the native tissue. For example, even state-of-the- art retinal differentiation protocols, which give rise to three dimensional retinal organoids, do not accurately recapitulate the cell-cell interactions that are present in the outer retina. Specifically, choroidal endothelial cells are rarely present and never form an organized layer beneath the RPE. Similarly, rather than forming a monolayer juxtaposed to newly developed photoreceptors, RPE cells are often found in clusters located at the edges of the neural retina. In this application we describe a program focused on the development and validation of a microphysiological system engineered to accurately recreate the complex architecture of the perfused outer retina. Specifically, we propose to combine ultra-high resolution, state-of-the-art 3D printing of retinal scaffolds, iPSC technology, and microfluidics to develop a next generation microphysiological system that recapitulates the outer retina unit. Completion of the aims outlined in this proposal will help pave the way for future studies focused on evaluating the pathophysiology of complex retinal degenerative disorders such as age-related macular degeneration and development of novel drug and gene-based therapeutics.