Development of a High Throughput Method for Vitrification

高通量玻璃化方法的开发

• 批准号: 10019053
• 负责人: LIA H CAMPBELL
• 金额: $ 27.53万
• 依托单位: TISSUE TESTING TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10019053
• 关键词: 3-Dimensional; 3D Print; Animal Model; Animals; Architecture; Automation; Back; Biological Assay; Biomedical Engineering; Blood Vessels; Cell Survival; Cells; Chemicals; Complex; Corrosion; Cosmetics; Cryopreservation; Cryopreserved Tissue; Custom; Development; Dimethyl Sulfoxide; Drug Modelings; Drug or Chemical; Drug or chemical Tissue Distribution; Ensure; Epithelial; Epithelium; Excision; Exposure to; Formulation; Future; Goals; Heart Valves; Human; Ice; In Vitro; Individual; Laboratories; Lead; Left; Legal patent; Logistics; Maintenance; Manufacturer Name; Measures; Methods; Modeling; Neck; Nitrogen; Outcome; Phase; Plastics; Polypropylenes; Polystyrenes; Process; Protocols documentation; Quality Control; Research; Rewarming; Robotics; Running; Sampling; Ships; Skin; Submersion; System; Temperature; Testing; Thinness; Time; Tissue Banks; Tissue Engineering; Tissue Model; Tissue Sample; Tissue Viability; Tissues; Toxic effect; Toxicity Tests; Toxicology; Variant; articular cartilage; base; cost; cryogenics; cytotoxicity; design; drug discovery; experimental study; human tissue; in vivo; innovation; irritation; outcome prediction; pre-clinical; predictive modeling; preservation; prototype; response; robotic system; screening; skin irritation; success; three-dimensional modeling; time use; tissue culture

ABSTRACT: In an effort to reduce the use of animals for toxicity testing, companies have developed bioengineered in vitro skin models and other human tissue constructs for predictive toxicity testing to replace traditional assays such as the Draize skin irritation test and also for use as preclinical animal models. Presently, tissues or skin constructs are made-to-order and require a lead time of several weeks before they can be used. Quality control checks are conducted post shipment creating serious recall liabilities for the manufacturer. The ability to cryopreserve tissue models would solve key logistical issues creating “on-demand” banks of tissues which can be prescreened to ensure quality control. This would create economies of manufacturing scale and reduce costs to manufacturers and end users. We have developed a patented ice-free vitrification method for cryopreservation of bioengineered skin constructs. Using this patented process, we have consistently demonstrated >80% viability of several types of human bioengineered epithelial constructs. These viability results persisted for several days post-rewarming and both viability and functional assessments meet the manufacturer's acceptance criteria for use in toxicology tests. However, a limiting factor for preservation at larger scale is the ability to vitrify multiple constructs at one time. Our current method allows for processing of up to 6-12 constructs at one time. Processing of more than 12 results in reduce viability due to cytotoxicity from exposure to cryoprotectants for too long. In this Phase I, proposal we plan to test the feasibility of using a specially designed cassette that can hold up to 24 constructs at once. The cassette would allow preservation of multiple constructs streamlining the vitrification process while removing variation between constructs due to operator error. The outcome will be measured by assessment of tissue viability and functional assays required by manufacturers for quality control. The success of this proposal will be inline with our company goal of commercializing our vitrification process by storing on-demand human bioengineered tissues for distribution to customers for drug or chemical screening and research applications. After demonstration of feasibility in Phase I, we will submit a Phase II proposal with the primary objective of custom development of a fully automated robotic sample handling system for construct cryopreservation and removal of cryoprotectants prior to use for predictive toxicity testing.

抽象的： 为了减少动物用于毒性测试的使用，公司在体外开发了生物工程 皮肤模型和其他人体组织结构用于预测毒性测试以取代传统测定 随着皮肤刺激性测试，并用作临床前动物模型。目前，组织或皮肤 构造是按订单进行的，需要花费数周的时间才能使用它们。质量控制 检查是在装运后进行的，为制造商制造了严重的召回责任。能力 冷冻水果组织模型将解决关键的后勤问题 预先筛选以确保质量控制。这将创造制造规模的经济并减少 制造商和最终用户的费用。我们已经开发了一种无冰的玻璃化方法 生物工程皮肤结构的冷冻保存。使用此专利过程，我们一直保持 几种类型的人类生物工程上皮构建体的可行性> 80％。这些可行性 甲型后几天的结果持续了几天，生存能力和功能评估都符合 制造商的接受标准用于毒理学测试。但是，保存的限制因素 较大的规模是一次染色多个构造的能力。我们当前的方法允许处理 一次最多6-12个结构。处理超过12个会导致由于细胞毒性引起的生存能力而导致的生存能力 暴露于冷冻保护剂太长。在此第I阶段，我们计划测试使用A的可行性 专门设计的盒式磁带，最多可容纳24个结构。盒子将允许保存 多种构造简化玻璃化过程，同时删除构造之间的变化 操作员错误。结果将通过评估组织生存能力和所需功能分析来衡量 由制造商进行质量控制。该提案的成功将与我们的公司目标隔离 通过存储按需人类生物工程组织以分配到分布到的玻璃化过程 药物或化学筛查和研究应用的客户。在阶段证明可行性之后 我，我们将提交一项II阶段提案，其主要目的是自动开发完全自动化 机器人样品处理系统用于构建冷冻保存和去除冷冻注视剂之前 预测毒性测试。