Biomimetic Materials and Bioprocesses for High-Density Cultures of Pancreatic Islets and Tissues

3.5 100万加拿大人患有糖尿病，相关费用为34亿美元。在1型和2型糖尿病中都观察到β细胞（产生胰岛素的细胞）丢失。胰岛移植可以恢复功能性β细胞，使1型糖尿病患者血糖正常。胰岛移植遇到了包括β细胞凋亡的问题。提高胰岛移植的成功率一直是研究界的目标。此外，由于缺乏足够的胰腺组织模型，制药公司已经确定治疗糖尿病的治疗剂的开发受到阻碍。 组织工程方法已经应用于在三维基质中包封和培养胰岛和β细胞，并使用生物反应器。这里的目的是将细胞/组织暴露在模拟其天然环境的环境中，因此它们可以表现得更具生理性。包裹胰岛可能是保护移植胰岛免受宿主反应的解决方案。组织工程的发展使得脱细胞器官和组织能够用作细胞治疗的支架。长期的研究目标是建立一种自动化的、高密度的胰腺组织和细胞的分离培养方法，并使其适用于其他组织和器官的培养，具体包括：1）建立一种灌注式生物反应器，用于器官/组织的脱细胞和细胞/组织的接种和培养。该反应器将用于研究灌注条件下脱细胞器官和组织中胰岛和β细胞的存活和功能;2）研究脱细胞组织/器官产生的仿生表面上内分泌胰岛/细胞的存活和功能;3）表征生物反应器灌注条件下内分泌胰岛和细胞与胰腺微血管的共培养。1）我们的仿生表面提供了优异的低污染涂层，允许通过消除非特异性细胞来研究特异性细胞反应。2)仿生支架如何影响胰岛的知识可以支持更好的封装技术的发展。3)了解流动条件如何影响胰岛反应，可以确定移植的胰岛如何受到植入部位生理条件的影响。开发一种自动化的去细胞化器官/组织的过程将是将手动操作的实验室规模方法转移到细胞治疗产品的工业过程的一个步骤。4)最后，关于如何在分离的胰岛中重建功能性脉管系统知之甚少。我们是第一个分离胰腺微血管的。该方案将支持高素质人员的培训。

3.5 million Canadians live with diabetes, with an associated cost of $ 3.4 billion. Beta-cells (cells producing insulin) lost has been observed in both types 1 and 2 diabetes. Pancreatic islet transplantation can restore functional beta-cells to achieve normoglycemia in type 1 diabetics. Islet transplantation has encountered problems including beta-cell apoptosis. Improving the success of islet transplantation has been a target in the research community. Also, the development of therapeutic to treat diabetes has been identified by pharmaceutical companies to be hindered due to a lack of adequate pancreatic tissue models. Tissue engineering methods have been applied to encapsulate and culture pancreatic islets and beta-cells in three-dimensional matrices, and using bioreactors. The aim here is to expose cells/tissues to environments mimicking their native ones, so they can behave more physiologically. Encapsulating pancreatic islets could be a solution to protect transplanted islets from host responses. Development in tissue engineering enables the use of decellularized organs and tissues as scaffolds for cell therapies. However, little is known about the mechanisms governing the effects of decellularized matrices on cells.The LONG-TERM PROGRAM OBJECTIVE is to develop the automated processes to isolate and culture endocrine pancreatic tissues and cells at high density and to adapt the process to other tissues and organs.The SPECIFIC OBJECTIVES are:1) To develop a perfusion bioreactor for organ/tissue decellularization and processes for cell/tissue seeding and culture. This reactor will be used to investigate pancreatic islets and beta-cells survival and function in decellularized organs and tissues under perfusion;2) To study endocrine pancreatic islets/cells survival and function on biomimetic surfaces produced from decellularized tissues/organs;3) To characterize the co-culture of endocrine pancreatic islets and cells with pancreatic microvessels under bioreactor perfusion.SIGNIFICANCE AND NOVELTY: 1) Our biomimetic surfaces provide excellent low-fouling coatings allowing studying specific cell responses by eliminating non-specific ones. 2) Knowledge about how biomimetic scaffolds affect islets can support the development of better encapsulation techniques. 3) Understanding how flow conditions affect islet responses allows identifying how transplanted islets are affected by physiological conditions at an implantation site. Developing an automated process to decellularize organs/tissues will be a step to transfer manually-operated laboratory-scale methods to industrial processes for cell therapy products. 4) Finally, little is known about how to re-establish a functional vasculature in isolated islets. We have been the first to isolate pancreatic microvessels. This program will support training of highly qualified personnel.