Combined Effects for Improved Tissue Dissociation

描述（申请人提供）：胰岛细胞移植为超过100万的I型（胰岛素依赖型）糖尿病患者提供了治愈的希望。然而，目前从供体胰腺回收胰岛细胞的方法留下了很大的改进空间。为了帮助改进这一程序，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证明一种处理因素的组合，这种组合应能显著改善目前将胰腺分离成可供移植的胰岛的方法。该小组建议通过结合三种处理方法来证明这一改进： 1.组织预处理（一致的薄切片）。可用于胶原酶消化的表面积和从组织表面到最内部细胞的距离都是影响酶对该过程的贡献的因素。组织预处理应最大限度地提高酶的有效性，同时最大限度地减少总细胞暴露时间。 2.增加能量。超声将用于加速解离过程。超声能量的空化作用将加速解离过程，并通过驱动局部混合来增强酶的作用。将评价超声的直接接触应用（使用探头）和间接应用（将组织浸入激发浴中）。 3.通风。通风将通过确保更好的氧气供应来保持细胞更有活力。适当的曝气还应通过提供播种气泡和减缓其崩溃速度来增加空化的有益效果。将对这些新工艺的单独和组合效应进行测试，并与辛辛那提目前使用的埃德蒙顿方案的当前计算机控制实施进行比较。

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.