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.

摘要 近三分之一的死者肝脏捐献者未用于移植或其他目的。许多器官 如果可以延长保存时间，则对于治疗和研究应用是有价值的。 超低温（< -140 ° C）冷冻保存可以实现无限期的器官储存。以前尝试 由于冰晶形成引起的细胞和结构破坏，器官冷冻保存失败。一 一种克服传统方法局限性的有前途的方法是玻璃化--即冷却 器官内的水不能经历从液态到固态冰的相变。 在冷冻保护剂（CPA）的帮助下，器官进入稳定的玻璃样状态，其中活的 理论上存储时间是无限的。然而，关键的挑战是在没有冰形成的情况下重新变暖， 开裂如果复温太慢，就会形成冰晶;如果复温不均匀，热应力就会导致开裂。 因此，升温的速度和均匀性至关重要。我们发明了一种新的复温方法 这就是所谓的“两个目标”。氧化铁纳米颗粒灌注在整个 沿着与CPA溶液接触的器官的脉管系统。然后可以通过冷却（现有的冷冻方法）将器官玻璃化。 技术），并根据需要通过将其放置在射频线圈中重新加热， 纳米颗粒，因此，从器官内。在初步研究中，我们第一次表明， 我们可以玻璃化和冷冻人类大小的（即猪）和大鼠肝脏，从而避免冰的形成或破裂。 并保持活力和器官水平的功能。在这一物理成功之后，我们在这里提出第一个 研究评估移植和生物活性/功能的成功，这些武装的器官。我们 中心假设是，通过玻璃化和冷冻保存将使功能性长期 用于移植、治疗和生物医学研究目的的全人肝脏库。虽然我们的长期 我们的目标是开发一种用于移植的冷冻保存人类肝脏的方法，我们对这个项目的目标是完善 全肝保存技术，以1）在保存期间改善保存的大鼠肝脏的体外和体内功能， 常温灌注和移植模型，2）确定细胞应激，损伤，和 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 大型动物（猪）和人类肝脏冷冻保存和复温。如果成功，这种方法可以 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。