Stasis technology for storage and transport of natural and engineered tissues

用于储存和运输天然组织和工程组织的静态技术

• 批准号: 9165581
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 22.59万
• 依托单位: TISSUE TESTING TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9165581
• 关键词: Allografting; Amputation; Animals; Antifreeze; Antifreeze Glycopeptides; Antifreeze Proteins; Apoptosis; Binding; Biological Assay; Biological Preservation; Blood Vessels; Calorimetry; Cartilage; Cell Death; Cell Survival; Cell physiology; Cells; Cessation of life; Chronic; Clinical; Complex; Cryopreservation; Cryopreserved Tissue; Development; Dry Ice; Dyes; Endothelial Cells; Engineering; Erythrocytes; Evaluation; Face; Feasibility Studies; Formulation; Freedom; Freezing; Glycerol; Glycolipids; Heart; Heart Valves; Hemolysis; Ice; Impairment; In Situ Nick-End Labeling; Insecta; Knee; Larynx; Lead; Leg; Licensing; Life; Limb structure; Literature; Liver; Malignant Neoplasms; Mammalian Cell; Measures; Methods; Organ; Organ Transplantation; Performance; Phase; Plants; Preparation; Procedures; Process; Protocols documentation; Recombinants; Recovery; Relaxation; Reporting; Research; Rewarming; Risk; Sampling; Services; Skeletal Muscle; Skin; Smooth Muscle Myocytes; Source; Staining method; Stains; Supplementation; System; Techniques; Technology; Testing; Tissue Banking; Tissue Banks; Tissue Engineering; Tissue Grafts; Tissue Model; Tissue Preservation; Tissue Sample; Tissue Viability; Tissues; Toxic effect; Trachea; Universities; Work; World Health Organization; abdominal wall; base; commercialization; cryobiology; cytotoxic; cytotoxicity; deep ocean; genus Solanum; improved; inhibitor/antagonist; innovation; interest; meetings; novel; prevent; professor; prospective; resazurin; scale up; small molecule libraries; technology development; teleost; therapy development

This proposal is focused on developing innovative new cryopreservation techniques employing ice-free preservation methods that will lead to the ability to bank complex vascularized tissue and organ grafts. Banking of large or complex tissues, such as limbs, and organs using current tissue banking practices employing conventional cryopreservation by freezing is not possible due to the well known damage caused by intra and extra-cellular ice formation. The company team has developed vitrification, cryopreserved storage in a “glassy” rather than crystalline ice phase, for banking of small tissue samples including blood vessels, heart valve tissues and cartilage. However, scale up to larger tissues, more complex tissues and organs needs better ice control during rewarming with less risk of cytotoxicity. Our collaborators, Professor Duman, University of Notre Dame, and Professor Ben, University of Ottawa, have developed natural and synthetic antifreeze compounds respectively that demonstrate benefits in frozen cell systems. Duman has developed recombinant insect- derived antifreeze proteins and isolation methods for antifreeze glycolipids from several insects and a plant that are very effective at ice control. Ben has developed a chemical library that was originally based upon an antifreeze glycoprotein derived from a deep sea teleost. We will evaluate the best of these compounds during ice-free cryopreservation using a vascular tissue model to determine if they improve ice control during rewarming and reduce the risk of cryoprotectant toxicity by allowing lower cryoprotectant concentrations to be used. There are three specific aims in this Phase I feasibility study to identify potentially marketable methods to increase efficacy and minimize tissue damage during cryopreservation of complex natural and engineered tissues: In Specific Aim 1 we will prepare of test compounds, antifreeze glycolipids from natural sources and synthesize large quantities of synthetic antifreeze compounds. A stock of recombinant insect-derived antifreeze proteins is already available. In Specific Aim 2 we will evaluate the impact of antifreeze compounds individually upon ice-free cryopreserved blood vessels and in Specific Aim 3 the best antifreeze supplements will be combined. Evaluations will include ice control during ice-free vitrification and tissue viability and function assays. The technology developed in this proposal will be equally effective for stasis, storage, of engineered tissue constructs as natural tissues. This work will lead to the development of banking, by us and other cryobiology groups, for internal organs, such as livers and hearts, vascularized composite allografts, such as larynx, trachea, abdominal wall, knee, skeletal muscles, facial tissues and legs, as well as improved methods for critical natural and engineered tissues such as skin, heart valves and blood vessels.

这项建议的重点是开发创新的新的冷冻保存技术，采用无冰 保存方法，这将导致银行复杂的血管化组织和器官移植的能力。银行 大的或复杂的组织，如四肢和器官，使用当前的组织库实践， 常规的冷冻低温保存是不可能的， 细胞外结冰该公司团队已开发出玻璃化，冷冻保存存储在一个“玻璃” 而不是结晶冰相，用于储存小组织样品，包括血管、心脏瓣膜 组织和软骨。然而，扩大到更大的组织，更复杂的组织和器官需要更好的冰 在复温过程中进行控制，细胞毒性风险较小。我们的合作者，杜曼教授，圣母大学 Dame和渥太华大学的Ben教授开发了天然和合成的防冻化合物 分别证明了在冷冻细胞系统中的益处。杜曼发明了重组昆虫- 衍生的抗冻蛋白和从几种昆虫和植物中分离抗冻糖脂的方法 在控制结冰方面非常有效。本开发了一个化学库，最初是基于一个 从深海硬骨鱼中提取的抗冻糖蛋白。我们将评估这些化合物中最好的， 使用血管组织模型进行无冰冷冻保存，以确定它们是否改善了冷冻过程中的冰控制 复温并通过允许较低的冷冻保护剂浓度来降低冷冻保护剂毒性的风险。 采用在第一阶段可行性研究中有三个具体目标，以确定潜在的可销售方法， 在冷冻保存复杂天然和工程化的 组织：在具体目标1中，我们将制备测试化合物，来自天然来源的防冻糖脂， 合成大量合成防冻剂化合物。重组昆虫源性 抗冻蛋白已经存在。在具体目标2中，我们将评估防冻剂化合物的影响 单独对无冰冷冻保存的血管和具体目标3最好的防冻补充剂 将被合并。评估将包括无冰玻璃化过程中的冰控制和组织活力和功能 测定。在这项建议中开发的技术将同样有效的停滞，存储，工程 组织构建体作为天然组织。这项工作将导致银行业的发展，由我们和其他 低温生物学组，用于内脏器官，如肝脏和心脏，血管化复合同种异体移植物，如 喉、气管、腹壁、膝、骨骼肌、面部组织和腿，以及改进的方法 用于关键的天然和工程组织，如皮肤、心脏瓣膜和血管。