Cryopreservation and nanowarming enables whole liver banking for transplantation, cell therapy and biomedical research

冷冻保存和纳米加温使整个肝脏库能够用于移植、细胞治疗和生物医学研究

• 批准号: 10584878
• 负责人: JOHN C BISCHOF
• 金额: $ 68.51万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10584878
• 关键词: Animals; Artificial Liver; Biocompatible Materials; Biological; Biomedical Research; Blood Vessels; Cell Therapy; Cellular Stress; Cessation of life; Chemicals; Cryopreservation; Cryoprotective Agents; Crystal Formation; Crystallization; Devices; Drug toxicity; Emergency Situation; Family suidae; Frequencies; Functional disorder; Glass; Goals; Heart; Heating; Hepatic; Hepatocyte transplantation; Human; Ice; In Vitro; Industry; Injury; Investigation; Ischemia; Kidney; Liquid substance; Liver; Lobe; Measures; Methods; Nanotechnology; Organ; Organ Preservation; Organ Size; Oryctolagus cuniculus; Osmosis; Pathway interactions; Patients; Performance; Perfusion; Pharmacologic Substance; Phase Transition; Predisposition; Preparation; Preservation Technique; Procedures; Protocols documentation; RF coil; Rattus; Recovery of Function; Research; Resource Allocation; Resources; Rewarming; Schedule; Science; Self-Help Devices; Solid; Specimen; Speed; Stress; System; Technology; Testing; Therapeutic; Therapeutic Research; Therapeutic Studies; Time; Toxic effect; Toxicity Tests; Transplantation; Transplantation Surgery; Water; cold temperature; cryogenics; disparity reduction; drug discovery; improved; in vivo; iron oxide nanoparticle; liver preservation; liver transplantation; nanoparticle; nanowarming; new technology; novel; organ allocation; pharmacologic; post-transplant; preservation; prevent; radio frequency; regenerative; scale up; success; thermal stress; transplant model

ABSTRACT Nearly one-third of deceased donor livers are unused for transplant or other purposes. Many of these organs would be valuable for therapeutic and research applications if preservation times could be extended. Cryopreservation at ultralow temperatures (< -140°C) can enable indefinite organ storage. Previous attempts at organ cryopreservation have failed due to cellular and structural disruption caused by ice crystal formation. One promising approach that overcomes the limitations of conventional strategies is vitrification – that is, cooling organs so quickly that the water within the organ cannot undergo the phase transition from liquid to solid ice. With the help of cryoprotective agents (CPAs), the organ enters a stable glass-like state wherein the viable storage time is theoretically unlimited. The critical challenge, however, is rewarming without ice formation or cracking. If rewarming is too slow, ice crystals form; if rewarming is not uniform, thermal stress causes cracking. Hence, speed and uniformity of warming are essential. We have developed a novel rewarming approach termed “nanowarming” that achieves both objectives. Iron oxide nanoparticles are perfused throughout the vasculature of the organ along with CPA solutions. The organ can then be vitrified by cooling (an existing technology) and rewarmed as needed by placing it in a radiofrequency coil that induces heating in the nanoparticles and, therefore, from within the organ. In preliminary studies, for the first time we have shown that we can vitrify and nanowarm human sized (i.e. porcine) and rat livers, thereby avoiding ice formation or cracking and preserving viability and organ-level function. Following on this physical success, we propose here the first study to assess both transplantation and biological viability/function success of these nanowarmed organs. Our central hypothesis is that cryopreservation by vitrification and nanowarming will enable functional long-term whole human liver banking for transplant, therapeutic and biomedical research purposes. While our long-term goal is to develop a method for cryopreserving human livers for transplant, our goals for this project are to refine whole-liver preservation technology to 1) improve in vitro and in vivo functionality of preserved rat livers during normothermic perfusion and in transplant models, 2) determine the mechanisms of cellular stress, injury, and death resulting from liver cryopreservation and strategies for injury mitigation, and 3) provide further investigation of large animal (porcine) and human liver cryopreservation and rewarming. If successful, this approach could revolutionize how these precious resources are allocated and utilized for patient and societal benefit.

抽象的 近三分之一的死亡捐献肝脏未用于移植或其他目的。 如果可以延长保存时间，对于治疗和研究应用将很有价值。 超低温（< -140°C）冷冻保存可以实现器官无限期保存。 由于冰晶形成造成细胞和结构破坏，器官冷冻保存失败。 克服传统策略局限性的有前景的方法是玻璃化——即冷却 器官的速度如此之快，以至于器官内的水无法经历从液态冰到固态冰的相变。 在冷冻保护剂（CPA）的帮助下，器官进入稳定的玻璃状状态，并且有活力 然而，储存时间理论上是无限的，但关键的挑战是在不形成冰或冰的情况下复温。 如果复温太慢，会形成冰晶；如果复温不均匀，热应力会导致开裂。 因此，升温的速度和均匀性至关重要。我们开发了一种新颖的复温方法。 被称为“纳米变暖”的技术可以实现这两个目标。 然后将器官的脉管系统与 CPA 溶液一起通过冷却（现有的方法）进行玻璃化。 技术），并根据需要将其放置在射频线圈中，在射频线圈中感应加热 纳米颗粒，因此，在初步研究中，我们首次证明了这一点。 我们可以对人类大小（即猪）和大鼠的肝脏进行玻璃化和纳米加热，从而避免结冰或破裂 继这一身体上的成功之后，我们在此提出第一个建议。 研究评估这些纳米加热器官的移植和生物活力/功能的成功。 中心假设是，通过玻璃化和纳米加热进行冷冻保存将使功能长期有效 整个人类肝脏库用于移植、治疗和生物医学研究目的。 目标是开发一种冷冻保存人类肝脏以进行移植的方法，我们这个项目的目标是完善 全肝保存技术1）改善保存的大鼠肝脏在体外和体内的功能 常温灌注和移植模型，2) 确定细胞应激、损伤和细胞应激的机制 肝脏冷冻保存导致的死亡和减轻损伤的策略，以及 3) 提供进一步的调查 如果成功的话，这种方法可以实现大型动物（猪）和人类肝脏的冷冻保存和复温。 彻底改变这些宝贵资源的分配和利用方式，以造福患者和社会。