Development of a High Throughput Method for Vitrification

高通量玻璃化方法的开发

基本信息

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
项目类别：
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

项目摘要

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：为了减少使用动物进行毒性测试，公司开发了体外生物工程用于预测毒性测试的皮肤模型和其他人体组织结构，以取代传统的检测方法，例如作为 Draize 皮肤刺激测试，目前也用作临床前动物模型、组织或皮肤。结构是按订单生产的，需要几周的交货时间才能使用质量控制。检查是在装运后进行的，这给制造商带来了严重的召回责任。冷冻保存组织模型将解决关键的后勤问题，创建“按需”组织库，可以进行预筛选以确保质量控制，这将创造制造规模经济并减少成本。我们开发了一种获得专利的无冰玻璃化冷冻方法。使用这种专利工艺，我们始终如一地冷冻保存生物工程皮肤结构。多种类型的人类生物工程上皮结构的活力已被证实超过 80%。结果在复温后持续数天，并且活力和功能评估均符合然而，制造商用于毒理学测试的验收标准是保存的限制因素。更大的规模是能够同时玻璃化多个结构，我们当前的方法允许处理。一次处理多达 6-12 个构建体，超过 12 个构建体会因细胞毒性而降低活力。暴露于冷冻保护剂的时间过长在第一阶段的提案中，我们计划测试使用冷冻保护剂的可行性。专门设计的盒式磁带可同时容纳多达 24 个构建体。多个结构简化了玻璃化过程，同时消除了结构之间的差异结果将通过评估组织活力和所需的功能测定来衡量。该提案的成功将符合我们公司的目标：通过存储按需人体生物工程组织并分发给我们，将我们的玻璃化过程商业化在阶段性论证可行性后，客户进行药物或化学品筛选和研究应用。 I，我们将提交第二阶段提案，其主要目标是定制开发全自动机器人样品处理系统，用于构建体冷冻保存并在使用前去除冷冻保护剂预测毒性测试。