Human Pancreatic Cells in Continuous Culture

连续培养的人胰腺细胞

• 批准号: 6587063
• 负责人: ROBERT J HAY
• 金额: $ 8.18万
• 依托单位: ROBERT H, LLC
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2004-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6587063
• 关键词: NOD mouse; RNA directed DNA polymerase; SCID mouse; cell line; enzyme linked immunosorbent assay; flow cytometry; gene expression; glucose; insulin; longevity; pancreatic islets; polymerase chain reaction; technology /technique development; telomerase; telomere; tissue /cell culture; transfection; western blottings

DESCRIPTION (provided by applicant): The proposed program as outlined should result in further important characterization and development of immortalized or restored cells of human pancreatic origin. Markers of differentiation and function, mainly insulin production and responsiveness to high glucose loads, will be used to identify populations of higher interest. These populations will be selected for transfection with the hTERT gene, and immortalized. Telomere-enhanced component strains will be isolated, identified, expanded, and further characterized. Telomerase activities and telomere length assays will be performed. To assess the functional consequences of the constitutive or periodic expression of hTERT, a series of structural and phenotypic tests will be performed. Positive controls will consist of related lines normal or immortalized in similar fashion but with the HPV16 E6/E7 genes (on hand). The functional integrity of our lines will be assessed in vivo, and our shielding strategy can be tested in NOD mice with autoimmune prone mechanisms in place. Thus, our cell line generation and engineering strategy should provide sets of potentially therapeutic populations for further developmental and transplantation research.

描述（由申请人提供）： 所提出的计划概述应导致进一步的重要表征和发展的永生化或恢复的人胰腺来源的细胞。分化和功能的标志物，主要是胰岛素的产生和对高葡萄糖负荷的反应，将用于识别更感兴趣的人群。将选择这些群体用于用hTERT基因转染，并使其永生化。端粒增强组分菌株将被分离、鉴定、扩增和进一步表征。将进行端粒酶活性和端粒长度测定。为了评估hTERT的组成性或周期性表达的功能后果，将进行一系列结构和表型测试。阳性对照将由正常或以类似方式永生化但具有HPV 16 E6/E7基因的相关细胞系组成（现有）。将在体内评估我们的细胞系的功能完整性，并且可以在具有自身免疫易感机制的NOD小鼠中测试我们的屏蔽策略。因此，我们的细胞系生成和工程策略应该提供一套潜在的治疗人群，为进一步的发展和移植研究。