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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.

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