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 将近三分之一的已故捐赠者生活未用于移植或其他目的。这些器官中有许多 如果可以延长准备时间，则对于治疗和研究应用是有价值的。 超高温下的冷冻保存（<-140°C）可以使无限期的器官存储。以前的尝试 器官冷冻保存由于冰晶形成引起的细胞和结构破坏而失败。一 有前途克服常规策略局限的有前途的方法是玻璃化 - 也就是说，冷却 器官如此之快，以至于器官中的水不能经历从液体到固体冰的相过渡。 在冷冻保护剂（CPA）的帮助下，器官进入一个稳定的玻璃状状态，其中可行 在理论上，存储时间是无限的。然而，关键的挑战是在没有冰形成或 破裂。如果转换太慢，则形成冰晶；如果转换不均匀，热应力会导致破裂。 因此，温暖的速度和均匀性至关重要。我们已经开发了一种新颖的恢复方法 称为“纳米武器”，实现了这两个目标。氧化铁纳米颗粒在整个过程中都被灌注 器官的脉管系统以及CPA溶液。然后可以通过冷却（现有的 技术），并根据需要将其放入诱导加热的射频线圈中。 纳米颗粒，因此从器官内部。在初步研究中，我们首次表明 我们可以染色和纳米臂大小（即猪）和老鼠的生活，从而避免冰形成或破裂 并保留生存能力和器官级功能。跟随这一身体上的成功，我们在这里提出了第一个 评估这些纳米臂器官的移植和生物生存能力/功能成功的研究。我们的 中心假设是，通过玻璃化和纳米武术的冷冻保存将使功能长期实现 整个人类肝脏银行，用于移植，治疗和生物医学研究目的。而我们的长期 目标是开发一种为移植人类生命而生命的方法，我们的目标是完善 全肝保存技术至1）改善保留老鼠生命的体外和体内功能 在移植模型中，2）确定细胞应激，损伤和 肝冷冻保存和减轻伤害策略造成的死亡，3）提供进一步的调查 大型动物（猪）和人肝冷冻保存和重新加热。如果成功，这种方法可以 彻底改变了这些宝贵的资源是如何分配和用于患者和社会福利的。