A perfluorocarbon-based culture device for beta cell biology applications (Phase

用于 β 细胞生物学应用的基于全氟化碳的培养装置（Phase

基本信息

批准号：
8487397
负责人：
Juan Dominguez-Bendala
金额：
$ 39.62万
依托单位：
OPHYSIO, INC.
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-10 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8487397
关键词：
Academia Address Administrator Adult Air Award Basic Science Beta Cell Biological Testing Biology Blood flow California Cell Culture Techniques Cell Death Cell Differentiation process Cell Survival Cell Therapy Cell physiology Cells Cellular biology Cessation of life Clinical Custom Development Devices Diabetes Mellitus Diffusion Digestive System Disorders Discipline Employee Strikes Ensure Equipment Evolution Fluorocarbons Funding Future Generations Goals Growth Human Hyperoxia Hypoxia Immunodeficient Mouse In Vitro Industry Institutes Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Islets of Langerhans Transplantation Kidney Diseases Knowledge Laboratories Legal patent Link Marketing Medical Membrane Methods Mission Modeling Organ Outcome Oxygen Oxygen measurement, partial pressure, arterial Pancreas Pattern Phase Physicians Physiological Production Public Health Regenerative Medicine Replacement Therapy Reproducibility Research Research Institute Silicones Speed Stem Cell Research Stem cells Sterilization Structure of beta Cell of islet System Technology Temperature Testing Tissues Translational Research Translations Transplantation Universities Washington Work Xenograft procedure base beta cell replacement cell type commercialization design diabetes mellitus therapy diabetic embryonic stem cell experience fetal flasks improved interest islet islet stem cells manufacturing process novel phase 1 study pre-clinical prevent prospective scale up stem cell differentiation stem cell therapy success technology development tissue culture tissue/cell culture tool trend

项目摘要

ABSTRACT Conventional culture vessels are not designed for physiological oxygen delivery. Both hyperoxia and hypoxia - commonly observed when culturing cells and tissues in regular plasticware- have been linked to reduced cellular function and death. An adequate means to provide oxygenation is also critical for stem cell applications in which the differentiation outcome is dependent on oxygen tension levels. We have addressed this problem by devising a novel culture device, the "oxygen sandwich". This simple system is designed to deliver oxygen in a quasi-physiological fashion by means of a basal air-permeable perfluorocarbon-silicone (PFC/Si) membrane. Our long-term goal is to establish this system as a new standard for tissue culture, both for applications in which oxygen diffusion rates become limiting (such as 3D culture) and for those that require precise adjustments of oxygenation to steer stem cell differentiation in the desired direction. We will focus our proof-of- concept work on islet/beta cell biology. This is a rapidly expanding market that includes clinical uses (islet transplantation), active in vitro research on adult/fetal islets of several species, and pre-clinicl stem cell research with the potential to revolutionize the treatment of diabetes within the next 5-10 years. The pertinence of our model choice is highlighted by two well-documented observations: [1] Pancreatic beta cells (the current end product used in clinical therapies for diabetes) are highly sensitive to sub- and super-physiological oxygen concentrations; and [2] Stem cell differentiation into beta cells (the subject of worldwide research to replace cadaveric islets for future clinical uses) is exquisitely dependent on evolving oxygen tensions. Our Phase I studies aimed at demonstrating that the enhanced in vitro survival and function observed in PFC/Si-cultured islets also resulted in better pre-clinical transplantation outcomes using a marginal mass xenotransplantation model (human islets into diabetic nu/nu immunodeficient mice). After the successful completion of these studies, our Phase II proposal is based on the following specific aims: (1) Scaling-up and definition of manufacturing process (QA & QC, sterilization) for mass production of PFC/Si culture devices; and (2) biological testing and reproducibility studies in human islets and embryonic stem cells (hESc). This application benefits from the assembly of first-rate teams with highly complementary expertise. Our Phase I results are strongly supportive of the feasibility of this proposal. Success in our research would fill a widely acknowledged gap in our ability to preserve islet cell function and survival in vitro confirming this system as a potential new standard for beta cell biology and differentiation studies. As such positive outcome might ultimately speed up the applicability of new-generation beta cell replacement therapies, this project is greatly relevant to the mission of the National Institutes of Diabetes and Digestive and Kidney Diseases (NIDDK). Success in these studies will also provide proof of principle of the superiority of our oxygen enhancing technology for many other cell culture applications.

抽象的常规培养容器不是为生理氧递送而设计的。在常规塑料软件中培养细胞和组织时通常观察到的高氧和缺氧 - 与细胞功能和死亡的降低有关。提供氧合的足够手段对于分化结果取决于氧气张力水平的干细胞应用也至关重要。我们通过设计一种新型的培养装置“氧气三明治”来解决了这个问题。这个简单的系统旨在通过基础可渗透的全氟苯甲酸（PFC/SI）膜以准生理学方式传递氧气。我们的长期目标是将该系统建立为组织培养的新标准，既适用于氧扩散速率成为限制的应用（例如3D培养），以及那些需要对氧合进行精确调整以使干细胞分化中的干细胞分化的应用。期望的方向。我们将把概念证明工作集中在胰岛/β细胞生物学上。这是一个迅速扩展的市场，包括临床用途（胰岛移植），几种物种的成人/胎儿胰岛的积极研究，以及临床前干细胞研究，有可能在接下来的5-10中彻底改变糖尿病的治疗年。我们的模型选择的相关性由两个有据可查的观察值强调：[1]胰腺β细胞（用于糖尿病临床疗法中使用的当前最终产物）对亚和超级生理氧浓度高度敏感； [2]干细胞分化为β细胞（全球研究的主题以替代尸体胰岛以供将来的临床用途）精致取决于不断发展的氧气张力。我们的I期研究旨在证明PFC/SI培养胰岛中观察到的增强的体外存活和功能也导致了使用边缘质量异种移植模型（人类胰岛中的糖尿病NU/NU/NU免疫缺陷小鼠）更好的临床前移植结果。这些研究成功完成后，我们的II期建议基于以下特定目的：（1）用于扩大和定义制造过程（QA＆QC，灭菌），用于PFC/SI培养设备的质量生产；（2）人类胰岛和胚胎干细胞（HESC）中的生物测试和可重复性研究。该应用程序受益于具有高度互补专业知识的一流团队的组装。我们的I阶段结果强烈支持该提案的可行性。在我们的研究中的成功将填补我们保留胰岛细胞功能和生存能力的广泛认可的差距，从而确认该系统是β细胞生物学和分化研究的潜在新标准。由于这种积极的结果最终可能会加快新一代β细胞替代疗法的适用性，因此该项目与美国国立糖尿病，消化和肾脏疾病（NIDDK）的任务非常相关。这些研究的成功还将为我们的氧气增强技术优越的原理证明，以实现许多其他细胞培养应用。