Bioluminescent Imaging of Pancreatic Islet Transplants

胰岛移植的生物发光成像

基本信息

批准号：
6786596
负责人：
Dixon B Kaufman
金额：
$ 31.62万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-09-30 至 2006-08-31
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): Our ultimate goal is to contribute in a meaningful way to the cure of diabetes through islet transplantation. We posit that early loss of transplant islet mass at the implantation site significantly reduces the functional potential of the freshly purified islet preparation. Certain anatomic and immunologic variables impact on the early fate of the implanted cells. The overall goal of the proposal is to define how the transplant location, the make-up of the transplanted cells (free cells, cellular matrix structures, or islets), and the immune responsiveness of the host affect cellular viability and function. We hypothesize that: 1) early loss of viable islet mass significantly reduces the functional potential of the freshly purified islet preparation; 2) that rates of loss of cell viability depend on engraftment site, the vigor of immediate host non-specific inflammatory responses, and the composition of the cellular structures transplanted. The specific aims are: 1. To develop a useful method of real-time bioluminescent imaging as a modality to monitor the fate of transplanted "tagged" islets from transgenic rat insulin II promoter (RIP)-luciferase donor strain mice. 2. To create effective adeno and adeno-associated viral vectors containing the RIP with two reporter genes: luciferase and green fluorescent protein for transfection of purified mouse and human islets. 3.To apply bioluminescent imaging techniques to monitor the fate of transplanted MIN6 ¿-cell lines expressing luciferase using a scaffold matrix. The aims will be accomplished by applying the general methods of real-time, non-invasive bioluminescent imaging to the specific field of islet transplantation. This imaging modality will permit visualization of "tagged' transplanted islets or ¿-cells in the living mouse. Through serial post-transplant monitoring, it will be possible to define how the fate of the viable islet mass is affected by anatomic and immunologic considerations that directly relate to transplant functional outcome.

描述（由申请人提供）：我们的最终目标是通过胰岛移植以有意义的方式为糖尿病的治疗做出贡献。我们认为植入部位移植胰岛质量的早期损失会显着降低新鲜纯化的胰岛制剂的功能潜力。某些解剖学和免疫学变量会影响植入细胞的早期命运。该提案的总体目标是确定移植位置、移植细胞的组成（游离细胞、细胞基质结构或胰岛）。和我们发现，宿主的免疫反应会影响细胞活力和功能：1）活胰岛质量的早期丧失会显着降低新鲜纯化的胰岛制剂的功能潜力；2）细胞活力丧失的速度取决于植入位点，直接宿主非特异性炎症反应的活力以及移植细胞结构的组成具体目标是： 1. 开发一种有用的实时生物发光成像方法作为监测移植细胞命运的方式。来自转基因大鼠胰岛素 II 启动子 (RIP)-荧光素酶供体品系小鼠的“标记”胰岛 2. 创建含有 RIP 的有效腺病毒和腺相关病毒载体，该载体具有两个报告基因：荧光素酶和绿色荧光蛋白，用于转染纯化的小鼠和小鼠。 3.应用生物发光成像技术监测移植的MIN6的命运。 -使用支架基质表达荧光素酶的细胞系将通过将实时、非侵入性生物发光成像的一般方法应用于胰岛移植的特定领域来实现。这种成像方式将允许“标记”的移植胰岛的可视化。通过连续移植后监测，可以确定与移植功能结果直接相关的解剖学和免疫学因素如何影响活胰岛团的命运。