Isochoric Pressure Based Preservation of Cells, Tissues and Organs

基于等容压的细胞、组织和器官保存

基本信息

批准号：
9910362
负责人：
Gerald Brandacher
金额：
$ 87.41万
依托单位：
SYLVATICA BIOTECH, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-08-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9910362
关键词：
3D Print Address Allografting Amputation Animal Model Animal Organ Animals Area Basic Science Behavioral Biocompatible Materials Biological Biological Preservation Biomechanics Biomedical Research Blood Vessel Tissue Blood Vessels Caenorhabditis elegans Cartilage Cell Survival Cell Therapy Cells Cessation of life Characteristics Chronic Clinical Clinical Protocols Communities Cryopreservation Cytoprotection Data Development Devices Digit structure Disease Emergency Situation Endothelium Ensure Equilibrium Evaluation Fertility Agents Fingers Fishes Food Forelimb Funding Agency Goals Government Graft Rejection Hand Heart Hour Human Ice In Vitro Letters Limb structure Measures Medical Research Metabolic Metabolism Methods Modeling Muscle Organ Organ Donor Organ Preservation Organ Transplantation Organism Ovary Patients Perfusion Phase Physical Function Process Protocols documentation Quality of life Rattus Readiness Recovery Recovery of Function Regenerative Medicine Research Research Personnel Rewarming Skin Skin Tissue Solid Structural Models System Temperature Testing Testis Thermodynamics Tissue Engineering Tissue Grafts Tissue Transplantation Tissue Viability Tissues Toxic effect Translations Transplant Recipients Transplantation Trauma Traumatic injury University of Wisconsin-lactobionate solution Upper Extremity Validation Work animation base biomaterial compatibility cell injury cell type clinical practice clinically relevant commercialization cryobiology cryogenics design drug discovery functional outcomes high throughput screening human model improved ischemic injury limb transplantation novel novel strategies oncofertility phase 2 study preclinical study preconditioning preservation pressure prototype reproductive organ scale up sensor stem stress tolerance success tissue/organ preservation transplant model trauma care

项目摘要

PROJECT SUMMARY/ ABSTRACT There is wide recognition within the transplant and broader biomedical communities, as well as government funding agencies, that extended and improved preservation of biological materials is needed for a huge range of endeavors in biomedicine, medical research and drug discovery, organ and tissue transplantation, cell-based therapies, fertility and regenerative medicine, emergency preparedness, and trauma care. The current standard of preservation for organs and vascular composite allografts (VCAs) consists of few hours of hypothermic static storage in UW solution on ice. This contributes to organ/VCA shortage and increased discard rates, exacerbates ischemic injury and graft rejection due to suboptimal donor-recipient matching, and diminishes the quality of life for transplant recipients. To address these challenges, we have pioneered a novel thermodynamic approach to biopreservation, based on a stable equilibrium state and isochoric (constant volume) condition, at high subzero (-5°C to -20 °C) temperatures, that will allow effective preservation of organs and VCAs, with a 20x-28x increase in storage duration over the current clinical practice (<6-12h of hypothermic storage) while avoiding cellular injury and other challenges created by storage at deep cryogenic temperatures. Importantly, we will build on the successful demonstration of feasibility in Phase I and the results that Sylvatica and UC Berkeley have collaboratively demonstrated with regard to high subzero isochoric preservation: (1) biocompatible thermodynamic pressure profiles for preservation cocktails that support human cell survival in isochoric systems with viabilities above 75%, (2) successful initial scale-up of isochoric preservation to whole rat hearts and skin at unprecedented temperatures (-8°C, -10°C), and (3) isochoric preservation of fish muscle and an entire organism (the research model C. elegans). Therefore, the objective of this Phase II proposal is to demonstrate, using animal and human models of VCA representative tissues and limb preservation, prolonged cryopreservation of VCAs for 5-7 days and weeks, or longer, with good functional outcome post storage and recovery. Across five specific aims, we will first employ skin and vascular models of VCAs for cryostasis cocktail and protocol optimization, with the central goal of enabling high subzero isochoric preservation while actively suppressing metabolism and enhancing stress tolerance. An isochoric preservation platform (chamber, pressure/temperature sensors) based on the successful system used in Phase I will be designed and built to support cryostasis protocols validation using clinical size human skin and blood vessels, then a model of rat forelimb preservation and assessment through pseudo transplantation, and then by orthotopic allotransplantation with comprehensive characterization of upper extremity functional and behavioral recovery. The results from the animal preservation/transplantation model will be used for the proof-of-concept for high subzero isochoric preservation of human fingers and hands, with full exsanguinous metabolic support quality assessment.

项目概要/摘要在移植界和更广泛的生物医学界以及政府中得到了广泛的认可资助机构认为，大量生物材料的保存需要延长和改进致力于生物医学、医学研究和药物发现、器官和组织移植、细胞基础治疗、生育和再生医学、应急准备和创伤护理的现行标准。器官和血管复合同种异体移植物 (VCA) 的保存包括几个小时的低温静态储存在冰上的 UW 溶液中这会导致器官/VCA 短缺并增加丢弃率，从而加剧这种情况。由于供体-受体匹配不佳而导致缺血性损伤和移植物排斥，并降低生活质量为了应对移植受者的这些挑战，我们开创了一种新颖的热力学。基于稳定平衡状态和等容（恒定体积）的生物保存方法在零度以下（-5°C 至 -20°C）的高温条件下，可以有效保存器官和 VCA，与当前临床实践相比，存储时间延长了 20 倍至 28 倍（<6-12 小时的低温储存），同时避免细胞损伤和深低温储存带来的其他挑战。重要的是，我们将在第一阶段成功论证可行性以及 Sylvatica 的结果的基础上和加州大学伯克利分校合作证明了零下高度等容保存：(1) 用于支持人体细胞等容生存的保存鸡尾酒的生物相容性热力学压力曲线系统活力高于 75%，(2) 成功地将等容保存初步扩大到整个大鼠心脏和皮肤在前所未有的温度（-8°C、-10°C）下，以及（3）鱼肌肉和整个生物体的等容保存（研究模型线虫）因此，第二阶段提案的目标是使用动物和模型进行演示。 VCA 人体模型的代表性组织和肢体保存，VCA 的长期冷冻保存 5-7 天和几周或更长时间，在存储和恢复后具有良好的功能结果，涵盖五个具体目标，我们将首先采用 VCA 的皮肤和血管模型进行冷冻鸡尾酒和方案优化，其中中央实现高零度以下等容保存的目标，同时积极抑制新陈代谢并增强基于压力耐受性的等容保存平台（腔室、压力/温度传感器）第一阶段使用的成功系统将被设计和构建，以支持使用以下方法进行低温冷冻协议验证：临床尺寸的人体皮肤和血管，然后通过以下方法建立大鼠前肢保存和评估模型假移植，然后通过具有上颌综合特征的原位同种异体移植动物保存/移植模型的结果将导致肢体功能和行为恢复。用于人类手指和手的高度零度以下等容保存的概念验证，进行全面的无血代谢支持质量评估。