PDF(Pubmed)
Abstract：
Pancreatic islet transplantation is one of the clinical options for certain types of diabetes. However, difficulty in maintaining islets prior to transplantation limits the clinical expansion of islet transplantations. Our study introduces a dynamic culture platform developed specifically for primary human islets by mimicking the physiological microenvironment, including tissue fluidics and extracellular matrix support. We engineered the dynamic culture system by incorporating our distinctive microwell-patterned porous collagen scaffolds for loading isolated human islets, enabling vertical medium flow through the scaffolds. The dynamic culture system featured four 12 mm diameter islet culture chambers, each capable of accommodating 500 islet equivalents (IEQ) per chamber. This configuration calculates > five-fold higher seeding density than the conventional islet culture in flasks prior to the clinical transplantations (442 vs 86 IEQ/cm2). We tested our culture platform with three separate batches of human islets isolated from deceased donors for an extended period of 2 weeks, exceeding the limits of conventional culture methods for preserving islet quality. Static cultures served as controls. The computational simulation revealed that the dynamic culture reduced the islet volume exposed to the lethal hypoxia (< 10 mmHg) to ~1/3 of the static culture. Dynamic culture ameliorated the morphological islet degradation in long-term culture and maintained islet viability, with reduced expressions of hypoxia markers. Furthermore, dynamic culture maintained the islet metabolism and insulin-secreting function over static culture in a long-term culture. Collectively, the physiological microenvironment-mimetic culture platform supported the viability and quality of isolated human islets at high-seeding density. Such a platform has a high potential for broad applications in cell therapies and tissue engineering, including extended islet culture prior to clinical islet transplantations and extended culture of stem cell-derived islets for maturation.
摘要：
胰岛移植是某些类型糖尿病的临床选择之一。然而,移植前难以维持胰岛限制了胰岛移植的临床扩展。我们的研究引入了一个动态培养平台，专门为初级人类胰岛开发，通过模仿生理微环境，包括组织流控和细胞外基质支持。我们通过整合我们独特的微孔图案化多孔胶原支架来设计动态培养系统，用于装载分离的人类胰岛，使垂直介质流过脚手架。动态培养系统具有四个12毫米直径的胰岛培养室，每个室能够容纳500个胰岛当量(IEQ)。该配置计算的接种密度比临床移植前的烧瓶中的常规胰岛培养物高>五倍(442对86IEQ/cm2)。我们用从已故捐献者中分离出的三批人类胰岛对我们的培养平台进行了为期2周的测试，超过了保持胰岛质量的常规培养方法的限制。静态文化作为对照。计算模拟显示，动态培养将暴露于致死缺氧（<10mmHg）的胰岛体积减少到静态培养的〜1/3。动态培养改善了长期培养中胰岛的形态退化，保持了胰岛的生存能力，缺氧标志物表达减少。此外，在长期培养中，动态培养保持了静态培养的胰岛代谢和胰岛素分泌功能。总的来说，生理微环境模拟培养平台支持高播种密度下分离的人类胰岛的生存能力和质量。这样的平台在细胞治疗和组织工程中具有广泛的应用潜力。包括在临床胰岛移植之前的延长胰岛培养和用于成熟的干细胞衍生胰岛的延长培养。
