Combined Effects for Improved Tissue Dissociation

描述(申请人提供)：胰岛细胞移植为100多万美国I型(胰岛素依赖型)糖尿病患者提供了治愈的希望。然而，目前从供体胰腺中回收胰岛细胞的方法仍有很大的改进空间。为了帮助改进这一过程，EnteraTech，Inc.与辛辛那提大学合作，正在开发一种新的胰岛细胞分离系统。第一阶段工作的总体目标将是展示一系列处理因素的组合，这应该会对目前将胰腺分离为可移植的胰岛的方法带来实质性的改进。该团队建议通过三种处理方法的组合来展示这一改进： 1.组织前处理(一致薄片)。胶原酶可用于消化的表面积和从组织表面到最里面细胞的距离都是影响酶对这一过程的贡献的因素。组织前处理应最大限度地提高酶的有效性，同时也最大限度地减少细胞的总暴露时间。 2.补充能量。超声波将被用来加速解离过程。超声能产生的空化作用会加速解离过程，并通过驱动局部混合来增强酶的作用。使用探头直接接触超声波应用，以及将组织浸入激发浴中的间接应用，都将被评估。 3.曝气。曝气将确保更好的氧气供应，从而使细胞更有活力。适当的曝气还应该通过提供播种气泡和减缓它们的破裂速度来增加空化的有益影响。将测试这些新工艺的单独效果和综合效果，并将其与辛辛那提大学目前使用的计算机控制的埃德蒙顿议定书的实施情况进行比较。

DESCRIPTION (provided by applicant): Islet cell transplantation offers the hope of a cure for the more than 1 million Americans who are type I (insulin dependant) diabetics. However, current methods of islet cell recovery from donor pancreata leave much room for improvement. To help improve this procedure, EnteraTech, Inc., in collaboration with The University of Cincinnati, is developing a novel islet cell dissociation system. The overall objective of the Phase I work will be to demonstrate a combination of processing factors which should bring substantial improvement to current methods for dissociating pancreata into viable islets for transplantation. The team proposes to demonstrate this improvement through the combination of three processing approaches: 1. Tissue preprocessing (consistent thin sectioning). Both the surface area available to the collagenase enzyme for digestion, and the distance from the tissue surface to the inner-most cell, are factors affecting the enzyme contribution to the process. Tissue preprocessing should maximize enzyme effectiveness while also minimizing total cell exposure time. 2. Addition of energy. Ultrasound will be used to accelerate the dissociation process. Cavitation from ultrasonic energy will accelerate the dissociation process and potentiate the action of the enzyme by driving local mixing. Both direct-contact application of ultrasound, with a probe, and indirect application, immersing the tissue in an excited bath, will be evaluated. 3. Aeration. Aeration will keep the cells more viable by assuring a better supply of oxygen. Appropriate aeration should also add to the beneficial effects of cavitation by providing seeding bubbles and moderating their speed of collapse. The individual and combined effects of these novel processes will be tested and compared against the current computer controlled implementation of the Edmonton Protocol currently in use at The University of Cincinnati.