Magnetic Separation of Liberated Islets During Isolation

分离过程中释放的胰岛的磁分离

• 批准号: 6931870
• 负责人: JEFFREY John CHALMERS
• 金额: $ 21.12万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931870
• 关键词: bioengineering /biomedical engineering; biomedical equipment development; flow cytometry; immunomagnetic separation; method development; pancreatic islet transplantation; pancreatic islets; protein quantitation /detection

DESCRIPTION (provided by applicant): This proposal focuses on the technical aspects of islet isolation, which remains a substantial impediment to implementation of islet transplantation as a treatment for type 1 diabetes. Removing islets as soon as they are liberated from the pancreatic tissue mass during isolation and circumventing the need for density gradient purification is likely to result in substantially increased islet yields and potencies from donor pancreases. Instant separation of liberated islets would minimize exposure to proteolytic enzymes, reactive oxygen intermediates, and mechanical stress associated with centrifugation. The hypervascularity of islets compared with acinar tissue allows their preferential enrichment with magnetic beads, which in turn allows their immediate separation in a magnetic field before overdigestion, fragmentation, and loss of islet viability ensues. This proposal seeks to exploit these principles in the development of magnetic separation-based techniques for porcine and human islet processing for the purpose of maximizing the utilization of donor pancreases for islet transplantation. The University of Minnesota Diabetes Institute for Immunology and Transplantation, the Ohio State University Department of Chemical Engineering and the Cleveland Clinic Foundation Department of Biomedical Engineering, have established an interdisciplinary partnership to address the following Specific Aims (SA). SA#1: To develop a magnetic flow sorter for continuous and instant separation of islets liberated during the islet isolation process. SA#2: To compare the yield and potency of porcine islets isolated using conventional versus novel, magnetic separation-based processing methods. SA#3: To examine the feasibility of instant magnetic separation of human islets. The successful completion of the proposed studies will supercede current islet isolation and purification techniques. This advance is expected to 1), increase threefold the number of patients rendered insulin independent from the available cadaver donor pool and 2), to permit treatment of patients with islets isolated from a hemi-pancreas from living donors.

描述(由申请人提供)：这项提案侧重于胰岛隔离的技术方面，这仍然是实施胰岛移植作为1型糖尿病治疗的一个重大障碍。在分离过程中，一旦胰岛从胰腺组织块中释放出来，立即将其移除，并避免密度梯度纯化的需要，可能会导致供体胰腺的胰岛产量和潜力显著增加。释放的胰岛的即时分离将最大限度地减少与离心相关的蛋白水解酶、活性氧中间体和机械应力的暴露。与腺泡组织相比，胰岛组织的多血管使其优先用磁珠浓缩，这反过来又允许它们在磁场中立即分离，以免随后发生过度消化、碎裂和胰岛活力丧失。这项建议旨在利用这些原则开发基于磁分离的猪和人胰岛加工技术，以最大限度地利用供体胰腺进行胰岛移植。明尼苏达大学糖尿病免疫和移植研究所、俄亥俄州立大学化学工程系和克利夫兰临床基金会生物医学工程系建立了跨学科合作伙伴关系，以实现以下具体目标(SA)。SA#1：开发一种磁流分选机，用于对在胰岛分离过程中释放的胰岛进行连续和即时的分离。 SA#2：比较使用传统和新的基于磁分离的处理方法分离的猪胰岛的产量和效力。SA#3：研究瞬时磁分离人类胰岛的可行性。拟议研究的成功完成将取代目前的胰岛分离和纯化技术。这一进展预计将1)，使胰岛素独立于现有身体捐赠者池的患者数量增加三倍；2)，允许治疗从活体捐赠者的半个胰腺中分离出来的胰岛患者。