Vanderlaan Emma L, Sexton Joshua, Evans-Molina Carmella, Buganza Tepole Adrian, Voytik-Harbin Sherry L
Weldon School of Biomedical Engineering, College of Engineering, Purdue University, West Lafayette, IN 47907, USA.
Medical Scientist/Engineer Training Program, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Lab Chip. 2023 Oct 10;23(20):4466-4482. doi: 10.1039/d3lc00371j.
The protection and interrogation of pancreatic β-cell health and function is a fundamental aspect of diabetes research, including mechanistic studies, evaluation of β-cell health modulators, and development and quality control of replacement β-cell populations. However, present-day islet culture formats, including traditional suspension culture as well as many recently developed microfluidic devices, suspend islets in a liquid microenvironment, disrupting mechanochemical signaling normally found and limiting β-cell viability and function . Herein, we present a novel three-dimensional (3D) microphysiological system (MPS) to extend islet health and function by incorporating a polymerizable collagen scaffold to restore biophysical support and islet-collagen mechanobiological cues. Informed by computational models of gas and molecular transport relevant to β-cell physiology, a MPS configuration was down-selected based on simulated oxygen and nutrient delivery to collagen-encapsulated islets, and 3D-printing was applied as a readily accessible, low-cost rapid prototyping method. Recreating critical aspects of the microenvironment within the MPS perfusion and islet-collagen interactions mitigated post-isolation ischemia and apoptosis in mouse islets over a 5-day period. In contrast, islets maintained in traditional suspension formats exhibited progressive hypoxic and apoptotic cores. Finally, dynamic glucose-stimulated insulin secretion measurements were performed on collagen-encapsulated mouse islets in the absence and presence of well-known chemical stressor thapsigargin using the MPS platform and compared to conventional protocols involving commercial perifusion machines. Overall, the MPS described here provides a user-friendly islet culture platform that not only supports long-term β-cell health and function but also enables multiparametric evaluations.
胰腺β细胞健康与功能的保护及研究是糖尿病研究的一个基本方面，包括机制研究、β细胞健康调节剂的评估以及替代β细胞群体的开发与质量控制。然而，当今的胰岛培养形式，包括传统的悬浮培养以及许多最近开发的微流控装置，都是将胰岛悬浮在液体微环境中，破坏了正常存在的机械化学信号，限制了β细胞的活力和功能。在此，我们提出了一种新型的三维（3D）微生理系统（MPS），通过结合可聚合的胶原蛋白支架来恢复生物物理支持和胰岛 - 胶原蛋白的机械生物学线索，从而延长胰岛的健康和功能。基于与β细胞生理学相关的气体和分子传输计算模型，根据模拟的氧气和营养物质向胶原蛋白包裹的胰岛的输送情况，选择了一种MPS配置，并将3D打印作为一种易于获取、低成本的快速成型方法。在MPS内重现微环境的关键方面，即灌注和胰岛 - 胶原蛋白相互作用，减轻了小鼠胰岛分离后的缺血和凋亡，为期5天。相比之下，以传统悬浮形式培养的胰岛则表现出逐渐缺氧和凋亡的核心区域。最后，使用MPS平台对胶原蛋白包裹的小鼠胰岛在有无著名化学应激剂毒胡萝卜素的情况下进行动态葡萄糖刺激胰岛素分泌测量，并与涉及商用灌注机的传统方案进行比较。总体而言，本文所述的MPS提供了一个用户友好的胰岛培养平台，不仅支持β细胞的长期健康和功能，还能进行多参数评估。
