A Novel Method to Isolate Islet Cells

分离胰岛细胞的新方法

• 批准号: 7272244
• 负责人: Jerry Knobeloch
• 金额: $ 18.12万
• 依托单位: ADVANCED BIOLOGICAL TECHNOLOGIES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-07 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7272244
• 关键词: Abattoirs; Acoustics; Address; Adoption; Affect; Agitation; Allogenic; Architecture; Area; Attention; Autologous; Beta Cell; Biological; Biological Assay; Biotechnology; Businesses; Capital; Cell Count; Cell Separation; Cell Survival; Cell Transplants; Cell physiology; Cells; Chemicals; Cities; Clinical; Collaborations; Commit; Computers; Condition; Deterioration; Development; Devices; Diabetes Mellitus; Digestion; Dissociation; Engineering; Ensure; Enzymes; Exclusion; Exposure to; Facility Construction Funding Category; Family suidae; Feasibility Studies; Fluorescent Dyes; Fracture; Frequencies; Funding; Goals; Grant; Growth; Head; Horns; Hospitals; Immunosuppression; Infusion procedures; Insulin; Insulin-Dependent Diabetes Mellitus; Intellectual Property; Investments; Islet Cell; Islets of Langerhans; Islets of Langerhans Transplantation; Laboratory Research; Lead; Length; Marble; Marketing; Measures; Mechanics; Methods; Mus; Numbers; Ohio; Operative Surgical Procedures; Organ Donor; Outcome; Oxygen; Pancreas; Pancreatectomy; Patients; Performance; Persons; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Physiologic pulse; Power Sources; Procedures; Process; Production; Pulse taking; Purpose; Rate; Reagent; Recovery; Regional Perfusion; Reporting; Research; Research Personnel; Review Committee; Solid; Source; Specimen; Staging; Standards of Weights and Measures; Stream; Structure of beta Cell of islet; Surface; Sus scrofa; System; Tail; Technology; Testing; Therapeutic immunosuppression; Thick; Time; Tissue Donors; Tissues; Transplant Recipients; Transplantation; Ultrasonics; Ultrasonography; United States National Institutes of Health; Universities; University Hospitals; Work; base; cell injury; chronic pancreatitis; commercialization; design; diabetic; exhaustion; experience; graft function; heat exchanger; improved; islet; nonhuman primate; novel; prevent; programs; prototype; response; skills; success; type I diabetic

DESCRIPTION (provided by applicant): The goals for this Phase I proposal are: 1) construction and bench testing of a novel device to dissociate tissue from donor pancreas using ultrasound for the recovery of pancreatic islet cells for transplantation, and 2) marked improvement in yields and viability of islets recovered using this approach as compared to the existing, standard method. Pancreatic islet cell transplantation has received considerable attention as a procedure with the potential to cure patients with diabetes. Allergenic transplantation as a potential cure for type 1 diabetes has been shown to be a realistic approach with several centers reporting insulin independence, or substantial improvement in patients who suffered from type I diabetes. In addition, auto-islet transplantation for persons undergoing pancreatectomy for chronic pancreatitis has been performed with increasing success. Despite these encouraging reports, several obstacles remain that prevent the widespread adoption of the application, including: The high level of skill required by those involved in the donor cell isolation and purification process. The lack of a clear understanding of the manner in which specific parameters of the isolation and purification process interact to affect outcomes and the inability to control these parameter, resulting in islet recovery rates at approximately 55 - 60% of viable cells. The severe shortage of donor organs. The need for long term immunosuppression and the deleterious effects of these drugs on long term islet cell function. The deterioration of graft function with time from islet exhaustion due to the transplantation of insufficient beta cell mass. It is our hypothesis that the current method of mechanical shaker agitation to dissociate pancreatic tissue is deleterious to optimum isolation results due to: 1) the need for a heat exchanger that results in a large thermal latency which exposes the cells to continued harmful enzymatic action while the system is cooled, 2) the inability of the shaker to dissociate the pancreas uniformly, and 3) the use of marbles to break apart the pancreas which results in physical damage at the cellular level. Advanced Biological Technologies, in collaboration with the University of Cincinnati, is developing a process to positively impact these obstacles through a novel use of ultrasonic technology. The system we describe herein is designed to fully exploit the spatial, temporal and thermal engineering parameters necessary to dissociate pancreatic tissue, thereby maximizing cell yield and viability in a consistent and repeatable manner while eliminating the need for a skilled operator. The objectives for the Phase I feasibility study are: Aim 1. Design and fabrication of sonic tissue dissociation system for the recovery of islets of Langerhans. Aim 2. Identify and assess initial performance profile that will be utilized for device optimization. Aim 3. Compare novel sonic approach to current islet cell dissociation method. Our team has more than 2 decades experience in device fabrication and testing for the isolation and perfusion of cells for research and transplantation and a clinical islet transplantation program with 75 patients transplanted in a five year period. Project Summary Successful application of this project would impact the quality and quantity of islet cells for transplantation into type I diabetics and for patients transplanted with islets to treat severe chronic pancreatitis. Specifically, the goal is to achieve insulin independence following a single islet infusion instead of the 2 - 3 donor pancreas per recipient currently needed in allergenic transplantation, thus allowing for more patients to receive this treatment. Additionally, and of no less importance, the integration of this technology with ABT's automated isolation device could be used to produce high quality islets for research purposes ensuring high quality islets on a consistent basis. Hence, the dissociation device is an enabling technology that would make it possible for hospitals, universities and companies involved with islet cell research to consistently obtain high quality islet cells for their work.

描述（由申请人提供）：该第一阶段提案的目标是：1）构建并进行台架测试一种新型装置，用于使用超声波从供体胰腺中分离组织，以恢复用于移植的胰岛细胞，以及 2）显着改善与现有的标准方法相比，使用这种方法回收的胰岛的产量和活力。胰岛细胞移植作为一种有望治愈糖尿病患者的手术受到了相当多的关注。变应原移植作为 1 型糖尿病的潜在治疗方法已被证明是一种现实的方法，多个中心报告称，I 型糖尿病患者的胰岛素独立性或显着改善。此外，对因慢性胰腺炎而接受胰腺切除术的患者进行自体胰岛移植也取得了越来越大的成功。尽管有这些令人鼓舞的报告，但仍然存在一些阻碍该应用广泛采用的障碍，包括： 参与供体细胞分离和纯化过程的人员需要高水平的技能。 对分离和纯化过程的特定参数相互作用影响结果的方式缺乏清晰的了解，并且无法控制这些参数，导致活细胞的胰岛回收率约为 55 - 60%。 供体器官严重短缺。 长期免疫抑制的需要以及这些药物对长期胰岛细胞功能的有害影响。 由于移植的β细胞量不足，导致胰岛衰竭，移植物功能随着时间的推移而恶化。我们的假设是，目前用机械摇床搅拌来分离胰腺组织的方法对于最佳分离结果是有害的，因为：1）需要热交换器，这会导致大的热潜伏期，使细胞暴露于持续有害的酶作用下，同时系统被冷却，2) 摇床无法均匀分离胰腺，3) 使用弹珠破碎胰腺，导致细胞水平的物理损伤。 Advanced Biological Technologies 与辛辛那提大学合作，正在开发一种通过超声波技术的新颖应用来积极影响这些障碍的工艺。我们在此描述的系统旨在充分利用解离胰腺组织所需的空间、时间和热工程参数，从而以一致和可重复的方式最大化细胞产量和活力，同时消除对熟练操作员的需求。第一阶段可行性研究的目标是： 目标 1. 设计和制造声波组织解离系统，用于朗格汉斯岛的恢复。目标 2. 确定并评估将用于设备优化的初始性能配置文件。目标 3. 将新颖的声波方法与当前的胰岛细胞解离方法进行比较。我们的团队在用于研究和移植的细胞分离和灌注的设备制造和测试以及在五年内移植了 75 名患者的临床胰岛移植项目方面拥有 20 多年的经验。项目概要 该项目的成功应用将影响移植到 I 型糖尿病患者以及移植胰岛治疗重症慢性胰腺炎的患者的胰岛细胞的质量和数量。具体来说，目标是在单次胰岛输注后实现胰岛素独立，而不是目前过敏性移植中每个受体需要 2 - 3 个供体胰腺，从而允许更多患者接受这种治疗。此外，同样重要的是，该技术与 ABT 自动隔离装置的集成可用于生产用于研究目的的高质量胰岛，从而确保始终如一的高质量胰岛。因此，解离装置是一项使能技术，使参与胰岛细胞研究的医院、大学和公司能够持续获得高质量的胰岛细胞用于其工作。