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.

项目总结/摘要 世界各地的实验室已经生产出数万种不同的斑马鱼品系 作为适用于人类遗传和生物医学研究的模式生物 遗传学和疾病维持所有这些有价值的基因型是昂贵的， 即使是最大的库存中心的容量。一种解决办法是冷冻保存斑马鱼 精子、卵子和胚胎，这种方法将使增加多- 使用NIH比较研究部设想的动物模型进行的机构研究 药斑马鱼的精子和卵子已经成功地冷冻保存，但技术 在最近的成功之前， 由明尼苏达大学的nanoComposix和John Bischof教授演示。的 斑马鱼胚胎冷冻保存的主要挑战是胚胎尺寸大，这限制了 胚胎可以在外部冷却和加热的速率。的创新办法 申请人使用的方法利用了金纳米颗粒的注射制剂， 高效加热器，当用红外激光照射时在内部产生热量。这 该技术首次展示了成功的冷却，低温稳定和 斑马鱼胚胎的复温进一步开发和商业化这一成功的 概念验证将解决斑马鱼研究人员的关键需求，并将为 在水产养殖和人类生殖方面的其他遗传保存应用， 满足社会、环境和生物多样性的需要。 本项目将进一步优化斑马鱼胚胎激光加温试剂、方案， 第一阶段合同期间开发的设备。该设备将进行beta测试， 斑马鱼研究中心和商业系统将被制造出来。系统将 由冷冻保护剂和金纳米颗粒组成的试剂配方组成， 优化以提供高光热转换效率和最小毒性，并且将 在符合ISO 13485的质量体系下生产。试剂微量注射到 胚胎和冷冻和激光复温方案将得到完善，以进一步改善胚胎 生存率，并将成为最终用户培训材料的基础。最后，第一代 设备将被开发，沿着测试自动化，高通量的方法， 胚胎冷却进一步的技术发展将提供通用的高通量 用于保存其他脊椎动物和非脊椎动物生物体的种质的平台。