Mechanical Induced Resonant Mixing for Islet Separation

用于胰岛分离的机械诱导共振混合

• 批准号: 6833722
• 负责人: FANGXIAO YANG
• 金额: $ 12.96万
• 依托单位: RESODYN CORPORATION
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6833722
• 关键词: athymic mouse; cell morphology; cell population study; cell sorting; collagenase; digital imaging; enzyme activity; morphometry; pancreatic islets; swine; technology /technique development

The proposed research is aimed to develop a mechanically-induced resonant mixing process for pancreas islet cell separation. The proposed study incorporates a unique mixing technology into a Resonant Enhanced Enzyme Tissue Digester (REETD) design. The REETD provides a novel method to enhance pancreas digestion and islet cell releasing processes without the use of foreign objects, such as marble or glass beads contacting the tissue. The technology provides enhanced mass transfer of digesting enzyme molecules into the pancreas and improves heat transfer of'a heated digesting solution'. These combined effects will greatly increase the tissue digestion rate and shorten the digesting time. Hence, both the islet cell recovery and their viability will be enhanced. Composition of the digestion enzyme solution will be optimized for fast release of islet cells, while preserving integrity of the islets. The post-digestion purification process will also be optimized and standardized by comparing various cell purification processes and methods. Because of the improved micromixing and mass transport properties that are created by the resonant mixing phenomena, the REETD technology is expected to: ? Increase enzymatic reaction efficiency ? Decrease digesting time ? Increase islet cell viability ? Improve islet recovery yield ? Preserve islets integrity ? Increase the separation reproducibility The proposed work is designed to establish a fundamental understanding of the governing mechanisms for applying mechanically-induced resonant mixing technology to reactor design aimed to improve the efficiency of enzymatic pancreas digestion and islet cell recovery process. This work meets the topic narrative for National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Division of Diabetes, Endocrinology and Metabolic Diseases - Islet Transplantation subtopic C "Development of methods to increase the yield and reproducibility of ....... " The proposed REETD technology development will be combined with enzyme composition optimization and, cell separation ant-purification methodology optimization. Commercially available collagenase and other digesting enzymes are ideal candidates for the study, but with some purification before their use. A salient feature of the resonant mixing technology is its capacity as compact and flexible tissue digestion reactor for a broad range of cell separation and purification processes.

本研究旨在开发一种机械诱导共振混合分离胰岛细胞的方法。拟议的研究将独特的混合技术纳入共振增强酶组织消化器（REETD）的设计。REETD提供了一种新的方法来增强胰腺消化和胰岛细胞释放过程，而不使用异物，如接触组织的大理石或玻璃珠。该技术提供了增强的消化酶分子进入胰腺的传质，并改善了“加热的消化溶液”的传热。这些综合作用将大大提高组织消化率，缩短消化时间。因此，胰岛细胞的恢复和它们的活力都将得到增强。消化酶溶液的组成将被优化以快速释放胰岛细胞，同时保持胰岛的完整性。消化后纯化工艺也将通过比较各种细胞纯化工艺和方法进行优化和标准化。由于共振混合现象产生的改进的微观混合和质量传输特性，REETD技术有望： ?提高酶促反应效率 ?缩短消化时间 ?增加胰岛细胞活力？提高胰岛回收率 ?保持胰岛完整性 ?提高分离重现性 本论文的目的是建立一个基本的理解机制，将机械诱导共振混合技术应用于反应器设计，旨在提高酶促胰腺的效率 消化和胰岛细胞回收过程。这项工作符合国家糖尿病、消化和肾脏疾病研究所（NIDDK）糖尿病、内分泌和代谢疾病分部-胰岛移植子主题C的主题叙述 “开发提高......产量和再现性的方法。" 建议REETD技术的发展将与酶组成优化和细胞分离纯化方法优化相结合。市售的胶原酶和其他消化酶是研究的理想候选物，但在使用前需要进行一些纯化。共振混合技术的一个显著特征是其作为紧凑和灵活的组织消化反应器的能力，用于广泛的细胞分离和纯化过程。