Magnetic Flow Sorter for Pancreatic Islet Isolation

用于胰岛分离的磁流分选机

• 批准号: 7861288
• 负责人: PAUL W TODD
• 金额: $ 6.83万
• 依托单位: TECHSHOT, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-30 至 2010-08-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7861288
• 关键词: Aliquot; Apoptosis; Architecture; Automation; Biological Assay; Blood Vessels; Cadaver; Caliber; Cell Size; Cell Survival; Cells; Characteristics; Clinic; Clinical; Clinical Trials; Collaborations; Computer software; DNA; Data; Dependence; Development; Devices; Diabetic mouse; Diagnosis; Digestion; Electronics; Engineering; Equipment; Exposure to; Family suidae; Foundations; Future; Gene Proteins; Genomics; Goals; Grant; Human; Human Resources; Indiana; Infusion procedures; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Islets of Langerhans; Islets of Langerhans Transplantation; Licensing; Magnetism; Marketing; Measurement; Measures; Medical Device; Metabolic; Methods; Microspheres; Minnesota; Modification; Molecular Profiling; Monitor; Mus; Nude Mice; Ohio; Operative Surgical Procedures; Organ; Organ Procurements; Oxygen Consumption; Pancreas; Particle Size; Patients; Performance; Phase; Phase I Clinical Trials; Polystyrenes; Positioning Attribute; Predisposition; Preparation; Procedures; Process; Proteomics; Protocols documentation; Public Health; Publications; Publishing; Pump; Quality Control; Reagent; Recovery; Regulation; Reporting; Research; Research Project Grants; Safety; Sampling; Site; Sorting - Cell Movement; Staining method; Stains; Stress; System; Techniques; Technology; Temperature; Test Result; Testing; TimeLine; Tissue Donors; Tissues; Transplantation; Universities; Work; Xenograft procedure; abstracting; base; biomaterial compatibility; cost; cytotoxicity; design; diabetic; diabetic patient; immunogenic; immunogenicity; innovation; instrumentation; islet; magnetic beads; magnetite ferrosoferric oxide; medical schools; meetings; minimally invasive; particle; pre-clinical; protein expression; prototype; research study; shear stress; success; type I diabetic; uptake; valve replacement

Project Summary/Abstract: SHOT, Inc., in collaboration with the University of Minnesota (UofM), Indiana University School of Medicine (IUSoM) and IKOtech LLC, proposes research leading to a magnetic separation technology that provides an improvement in potency and yield of pancreatic islets for transplantation. Islet transplantation is a method of treatment that has successfully reversed insulin dependence in Type 1 diabetics in clinical trials. However, one of the most significant issues with bringing pancreatic islet transplantation to the greater public is the difficulty of isolating a critical mass of healthy, viable islets from a single donor pancreas to achieve a successful clinical transplant. Current isolation and purification techniques lose 30-50% of the original islet mass and result in cellular apoptosis and/or loss of viability. This loss of viable islets almost always results in the need to utilize two pancreases for a single transplant (the "Edmonton Protocol"). The proposed innovation is a revolutionary device that is capable of magnetically isolating pancreatic islets with the goal of successful single-organ islet transplantations. The ultimate goal of this research is to increase the quality of healthy, viable, isolated, and purified islets recovered during the isolation process yielding purified islets ideally suited for transplantation into Type I diabetic patients. This new purification system is based on Quadrupole Magnetic flow Sorting (QMS) technology, which has been licensed by IKOtech from The Ohio State University and Cleveland Clinic Foundation. Available QMS hardware is capable of separating cell-sized particles (6-40 ¿m diameter); however, processing the larger islets (150-350 ¿m) requires significant modifications to the QMS hardware. During Phase I research SHOT successfully developed the key technologies required for the processing of intact islets by QMS. The new QMS will solve some of the well- known problems associated with human and xenogeneic islet preparations. For this proposed Phase II project, SHOT will (1) complete the design and manufacture of a QMS system capable of isolating pancreatic islets, (2) in conjunction with external colleagues, research and identify available magnetic reagents with a particle size and magnetic susceptibility that is optimized for isolating islets in three porcine isolation trials and that meets appropriate biocompatibility (i.e. cytotoxicity and immunogenicity) requirements, and (3) verify the performance of the technology by testing the QMS system and reagents together in three standardized porcine pancreatic islet isolations in collaboration with the islet isolation team at UofM. Islet isolation quality will be assessed via islet count, Oxygen Consumption Rate (OCR) and OCR per mg of DNA (OCR/DNA) measurements, proteomic and genomic molecular profiling, and diabetic nude mouse bioassays. The Phase II effort is designed to prove the reasonable safety and efficacy of the QMS technology for isolating pancreatic islets and will result in preclinical data in support of a Phase II Competing Continuation grant for required testing, eventually leading to an Investigational Device Exemption (IDE) approving QMS for clinical trials. Project Narrative: The project ¿Magnetic Flow Sorter for Pancreatic Islet Isolations¿ is relevant to public health as an enabling technology for pancreatic islet transplantation, which is a promising and minimally invasive cure for Type 1 Diabetes currently undergoing clinical trials. Type 1 Diabetes currently afflicts an estimated 800,000+ nationally with 60,000 or more cases diagnosed each year. Severe organ procurement limitations currently restrict the number of patients that can be treated via islet transplantation (which is compounded by the need for more than one organ per transplant), so success with the proposed technology will double the number of patients that can be cured with islets transplanted from cadaver donor tissue and will potentially facilitate unlimited treatment with the advent of porcine xenotransplantation as a further enabling technology.

项目摘要/摘要：SHOT, Inc. 与印第安纳州明尼苏达大学 (UofM) 合作 大学医学院 (IUSoM) 和 IKOtech LLC 提出了磁分离研究 提高胰岛移植效力和产量的技术。胰岛 移植是一种成功逆转 1 型糖尿病患者胰岛素依赖的治疗方法 在临床试验中。然而，将胰岛移植引入临床的最重要问题之一是 更广泛的公众认识到，从单个供体胰腺中分离出足够数量的健康、有活力的胰岛是很困难的。 取得成功的临床移植。目前的分离和纯化技术损失了 30-50% 原始胰岛质量并导致细胞凋亡和/或活力丧失。存活胰岛的损失几乎 总是导致需要利用两个胰腺进行单次移植（“埃德蒙顿方案”）。这 所提出的创新是一种革命性的装置，能够通过磁力隔离胰岛 成功的单器官胰岛移植的目标。这项研究的最终目标是提高 在分离过程中回收的健康、有活力、分离和纯化的胰岛质量，产生纯化的胰岛 胰岛非常适合移植到 I 型糖尿病患者体内。这种新的净化系统基于 四极磁流分选（QMS）技术，已获得俄亥俄州IKOtech授权 州立大学和克利夫兰诊所基金会。可用的 QMS 硬件能够分离细胞大小的 颗粒（直径 6-40 µm）；然而，处理较大的小岛（150-350 m）需要大量的 对 QMS 硬件的修改。在第一阶段研究期间，SHOT 成功开发了关键 QMS 处理完整胰岛所需的技术。新的质量管理体系将解决一些问题 与人类和异种胰岛制剂相关的已知问题。对于这个拟议的二期项目， SHOT 将 (1) 完成能够分离胰岛的 QMS 系统的设计和制造，(2) 与外部同事合作，研究并确定可用的磁性试剂的粒径 和磁化率，在三项猪分离试验中针对分离胰岛进行了优化，并且满足 适当的生物相容性（即细胞毒性和免疫原性）要求，以及（3）验证性能 通过在三个标准化猪胰腺中一起测试 QMS 系统和试剂来验证该技术 与密歇根大学的胰岛隔离团队合作进行胰岛隔离。胰岛隔离质量将通过以下方式评估 胰岛计数、耗氧率 (OCR) 和每毫克 DNA 的 OCR (OCR/DNA) 测量、蛋白质组学 和基因组分子分析以及糖尿病裸鼠生物测定。第二阶段的努力旨在证明 QMS 技术用于分离胰岛的合理安全性和有效性，并将导致 临床前数据支持所需测试的第二阶段竞争继续资助，最终导致 研究设备豁免 (IDE) 批准临床试验的质量管理体系。项目叙述：项目“用于胰岛隔离的磁流分选机”是相关的 作为胰岛移植的一项使能技术，对公众健康产生了重大影响，这是一项 目前正在进行临床试验，有望以微创方式治愈 1 型糖尿病。 目前，全国约有 800,000 多人患有 1 型糖尿病，其中 60,000 例或更多病例 每年都会确诊。目前，严格的器官获取限制限制了器官获取的数量。 可以通过胰岛移植进行治疗的患者（由于需要更多的胰岛移植而变得更加复杂） 每次移植一个器官），因此所提出的技术的成功将使数量增加一倍 可以通过从尸体供体组织移植的胰岛来治愈的患者，并将 随着猪异种移植作为一种治疗方法的出现，可能会促进无限的治疗 进一步的赋能技术。