Mechanical Induced Resonant Mixing for Islet Separation

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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.