A study of Model Beta-cells in Diabetes Treatment

模型β细胞在糖尿病治疗中的研究

• 批准号: 6728267
• 负责人: IOANNIS CONSTANTINIDIS
• 金额: $ 26.19万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-03-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6728267
• 关键词: Krebs' cycle; Langerhans' cell; NOD mouse; acidity /alkalinity; alginates; artificial endocrine pancreas; bioenergetics; blood glucose; diabetes mellitus therapy; disease /disorder model; glucose metabolism; glycolysis; hypoxia; insulin; medical implant science; nonhuman therapy evaluation; nuclear magnetic resonance spectroscopy; pancreatic islets; peritoneal cavity; phosphorylation; polylysine; secretion; swine; tissue /cell culture; tissue engineering

Description (provided by applicant): The objective of the proposed research is to investigate the significance of cellular bioenergetics to insulin secretion in a bioartificial pancreatic construct composed of transformed beta-cells under environmental conditions that prevail in vivo at the site of implantation. Since transformed beta-cell lines are proposed as alternatives to islets in the development of a bioartificial pancreas, it is imperative that we understand how insulin secretion is affected at these sites. It is our hypothesis that transformed cell lines are more tolerant than mammalian islets to environmental conditions experienced in the peritoneal cavity, a site commonly used for the implantation of these constructs. Experiments proposed in this application will be performed both in vitro and in vivo. Specifically, in vitro experiments will focus on the quantification of glucose metabolism and resultant phosphorylation potential for bioartificial pancreatic constructs composed of betaTC-tet, and INS-1(832/13) cells, as well as porcine islets encapsulated in alginate/poly-L-lysine/alginate (APA) beads as a function of oxygen concentration and pH. Glucose metabolism will be assessed via 13C NMR spectroscopy by measuring the flux through glycolysis, citric acid cycle, and key anaplerotic pathways, while the phosphorylation potential will be quantified via 31P NMR spectroscopy. In vivo experiments will focus on a device that will contain the APA beads so that they are not dispersed throughout the peritoneal cavity and thus NMR experiments performed in vitro can also be performed in vivo. Preliminary data presented in this revised application support our hypothesis and provide the foundation to ensure that proposed experiments are feasible and can be completed successfully. Overall, we believe that the acquired data will contribute significantly to our understanding of the mechanism of insulin secretion, as well as extend our knowledge in the field of tissue engineering.

描述（由申请人提供）：拟议研究的目的是 研究细胞生物能量对胰岛素分泌的重要性 在由转化的β细胞组成的生物人工胰腺结构中 在环境条件下，在体内盛行的现场 植入。由于提出了转换的β电池线作为替代方案 在生物人工胰腺发展中的胰岛，我们必须必须 了解在这些部位如何影响胰岛素分泌。这是我们的 转化的细胞系比哺乳动物胰岛更耐受的假设 对于腹膜腔体经历的环境条件 通常用于植入这些结构。 在本应用中提出的实验将在体外和 体内。具体而言，体外实验将集中于定量 葡萄糖代谢和生物人造的磷酸化潜力 胰腺构建体由BETATC-TET和INS-1（832/13）细胞组成 作为猪胰岛封装在藻酸盐/poly-L-赖氨酸/藻酸盐（APA）珠中 作为氧浓度和pH的函数。葡萄糖代谢将是 通过通过糖酵解测量通量，通过13C NMR光谱评估 柠檬酸循环和关键的消毒途径，而磷酸化 电势将通过31p NMR光谱进行定量。体内实验将 专注于将包含APA珠的设备，以免它们 分散在腹膜腔中，因此进行了NMR实验 体外也可以在体内进行体外。在此提供的初步数据 修订的申请支持我们的假设，并为确保 提出的实验是可行的，可以成功完成。 总体而言，我们认为获取的数据将对我们的 了解胰岛素分泌机制，并扩展我们 在组织工程领域的知识。