A perfluorocarbon-based culture device for beta cell biology applications

说明（由申请人提供）：传统的培养容器不是为生理氧输送而设计的。在普通塑料器皿中培养细胞和组织时经常观察到的高氧和低氧都与细胞功能下降和死亡有关。胰腺细胞尤其如此，它们对亚生理氧浓度和超生理氧浓度高度敏感。为了解决这个问题，我们设计了一种新的培养装置——“氧气三明治”。这个简单的系统被设计成通过一个基本的透气性全氟碳硅（PFC/Si）膜以一种类似生理的方式输送氧气。我们的长期目标是将该系统建立为β细胞生物学应用的新培养标准，这是一个迅速扩大的市场，包括任何物种的成人/胎儿胰岛的体外研究，以及β细胞分化研究。在此背景下，本I期研究的具体目的是建立概念证明，PFC/ si培养胰岛的体外存活和功能增强导致更好的临床前移植结果。我们的研究设计基于边际质量异种移植模型（人类胰岛移植到糖尿病nu/nu免疫缺陷小鼠中），在移植前进行PFC/Si培养的潜在益处应该转化为与对照（常规培养）胰岛移植的动物相比，糖尿病逆转的时间更短。我们的初步结果强烈支持这一建议的可行性，我们已经与迈阿密大学糖尿病研究所合作，以及时实现我们的目标。我们的研究成功将填补我们在体外保护胰岛细胞功能和存活能力方面广泛承认的空白，确认该系统作为β细胞生物学和分化研究的潜在新标准。这一积极结果将与美国国家糖尿病、消化和肾脏疾病研究所（NIDDK）的使命高度相关，并可能最终对人类胰岛/ β细胞移植具有临床意义。公共卫生相关性：β细胞研究是一个很大的领域，目前（胰岛移植）和未来（干细胞和异种移植）的糖尿病临床治疗进一步推动了基础科学的兴趣。然而，由于传统培养系统无法提供生理氧合，β细胞研究受到损害。由于我们的提案旨在测试一种专门设计用于克服这一问题的新型培养装置，因此这些研究与国家糖尿病、消化和肾脏疾病研究所（NIDDK）的使命高度相关，并可能最终对公众健康产生重大影响。

DESCRIPTION (provided by applicant): Conventional culture vessels are not designed for physiological oxygen delivery. Both hyperoxia and hypoxia -commonly observed when culturing cells and tissues in regular plastic ware- have been linked to reduced cellular function and death. This is particularly true of pancreatic beta cells, which are highly sensitive to sub- and super-physiological oxygen concentrations. We have addressed this problem by devising a novel culture device, the "oxygen sandwich". This simple system is designed to deliver oxygen in a physiological-like fashion by means of a basal air-permeable perfluorocarbon-silicone (PFC/Si) membrane. Our long-term goal is to establish this system as the new culture standard for beta cell biology applications, a rapidly expanding market that includes in vitro research on adult/fetal islets of any species, as well as beta cell differentiation studies. In this context, the specific aim of this Phase I proposal is to establish proof of concept that the enhanced in vitro survival and function observed in PFC/Si-cultured islets results in better pre-clinical transplantation outcomes. Our research design is based on a marginal mass xenotransplantation model (human islets into diabetic nu/nu immunodeficient mice), where the potential benefits of PFC/Si culture prior to transplantation should translate into a shorter time to diabetes reversal compared to animals transplanted with control (regular culture) islets. Our initial results are strongly supportive of the feasibility of this proposal, and we have partnered with the Diabetes Research Institute at the University of Miami to accomplish our goals in a timely manner. 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. Such positive outcome would be highly relevant to the mission of the National Institutes of Diabetes and Digestive and Kidney Diseases (NIDDK), and might ultimately be of clinical significance for human islet/beta cell transplantation. PUBLIC HEALTH RELEVANCE: Beta cell research is a large field where basic science interest is further fueled by current (islet transplantation) and prospective (stem cells and xenotransplantation) clinical therapies for diabetes. However, beta cell studies are compromised by the inability of conventional culture systems to provide physiological oxygenation. Since our proposal aims at testing a novel culture device specifically designed to overcome this problem, these studies are highly relevant to the mission of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) and may ultimately have a significant impact on public health.