Gold nanoparticle laser warming of cryopreserved zebrafish embryos

金纳米颗粒激光对冷冻斑马鱼胚胎的加温

• 批准号: 10016844
• 负责人: JOHN C BISCHOF
• 金额: $ 72.81万
• 依托单位: NANOCOMPOSIX, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10016844
• 关键词: 3D Print; Address; Adult; Animal Model; Aquaculture; Biodistribution; Biodiversity; Biomedical Research; Chorion; Contracts; Cryopreservation; Crystallization; Development; Dimensions; Diploidy; Disease; Embryo; Equipment; Feasibility Studies; Fishes; Formulation; Freezing; Generations; Genes; Genetic; Genetic Diseases; Genetic Research; Genome; Genotype; Goals; Health; Hour; Human; Human Genetics; Ice; International; Laboratories; Lasers; Legal patent; Letters; Malignant Neoplasms; Measures; Medicine; Methods; Microinjections; Minnesota; Modeling; Organ; Organism; Other Genetics; Performance; Pharmacologic Substance; Phase; Physiologic pulse; Polyethylene Glycols; Process; Protocols documentation; Publishing; Reagent; Reproduction; Research; Research Personnel; Rewarming; Surface; Survival Rate; System; Techniques; Technology; Testing; Time; Tissue Preservation; Toxic effect; Training; United States National Institutes of Health; Universities; Work; Zebrafish; assisted reproduction; base; biomaterial compatibility; commercial application; commercialization; comparative; cryogenics; efficacy testing; egg; embryo cryopreservation; embryo preservation; improved; innovation; interest; nanoGold; nanoparticle; preservation; prevent; reproductive; sperm cell; success; technology development; trait; zebrafish genome

Project Summary/Abstract Laboratories around the world have produced tens of thousands of distinct zebrafish lines that serve as model organisms for genetic and biomedical research that applies to human genetics and diseases. Maintaining all these valuable genotypes is expensive and beyond the capacity of even the largest stock centers. One solution is the cryopreservation of zebrafish sperm, eggs and embryos, an approach that would enable the strategy of increased multi- institutional research using animal models envisioned by the NIH Division of Comparative Medicine. Zebrafish sperm and eggs have been successfully cryopreserved, but technical challenges had prevented the cryopreservation of zebrafish embryos until recent success demonstrated by nanoComposix and Prof. John Bischof at the University of Minnesota. The main challenge with zebrafish embryo cryopreservation is the large embryo size, which limits the rate at which the embryo can be externally cooled and warmed. The innovative approach used by the applicants utilizes an injected formulation of gold nanoparticles that act as ultra- efficient heaters to generate heat internally when illuminated with an infrared laser. This technology demonstrated, for the first time, the successful cooling, cryogenic stabilization and rewarming of zebrafish embryos. Further development and commercialization of this successful proof-of-concept will address a critical need for zebrafish researchers and will form the basis for other genetic preservation applications in aquaculture and human reproduction, improving reproduction for societal, environmental and biodiversity needs. This project will further optimize the zebrafish embryo laser warming reagents, protocols, and equipment developed during the Phase I contract. The equipment will be beta tested by zebrafish research centers and a commercial system will be manufactured. The system will consist of a reagent formulation composed of a cryoprotectant agent and a gold nanoparticle, optimized to provide high photothermal conversion efficiency and minimal toxicity, and will be manufactured under a ISO 13485-compliant quality system. Reagent microinjection into the embryos and freezing and laser rewarming protocols will be refined to further improve embryo survival rates and will form the basis of training materials for end users. Finally, first-generation equipment will be developed, along with testing of automated, high-throughput methods for embryo cooling. Further technology development will provide a universal high throughput platform for preserving germplasm of other vertebrate and non-vertebrate organisms.

项目摘要/摘要 世界各地的实验室都产生了成千上万个不同的斑马鱼线 作为适用于人类的遗传和生物医学研究的模型生物 遗传学和疾病。维护所有这些有价值的基因型是昂贵的，超出了 甚至最大的股票中心的容量。一种解决方案是斑马鱼的冷冻保存 精子，鸡蛋和胚胎，一种方法，可以使多种多样的策略 NIH比较分区设想的动物模型的机构研究 药品。斑马鱼精子和鸡蛋已经成功保存了，但是技术 挑战阻止了斑马鱼胚胎的冷冻保存，直到最近成功 明尼苏达大学的Nanocomposix和John Bischof教授展示。这 斑马鱼胚胎冷冻保存的主要挑战是较大的胚胎大小，它限制 胚胎可以在外部冷却和温暖的速率。创新的方法 申请人使用的是用注入的金纳米颗粒的配方 用红外激光照亮时，有效的加热器可以在内部产生热量。这 技术首次证明了成功的冷却，低温稳定和 斑马鱼胚胎的重新加热。这项成功的进一步发展和商业化 概念证明将满足斑马鱼研究人员的迫切需求，并将构成 在水产养殖和人类繁殖中的其他遗传保存应用，改善 社会，环境和生物多样性需求的繁殖。 该项目将进一步优化斑马鱼激光变暖试剂，协议， 和在第一阶段合同期间开发的设备。设备将通过Beta测试 斑马鱼研究中心和商业系统将被制造。系统将 由由冷冻保护剂和金纳米颗粒组成的试剂制剂组成， 优化以提供高光热转化效率和最小的毒性，将是 在ISO 13485兼容的质量系统下制造。试剂微注射 胚胎和冷冻和激光重新加热方案将得到改进，以进一步改善胚胎 生存率将构成最终用户培训材料的基础。最后，第一代 设备将开发，以及测试自动化的高通量方法 胚胎冷却。进一步的技术开发将提供通用的高通量 保留其他脊椎动物和非脊椎动物生殖种质的平台。