Development of Germplasm Resources for Preservation of Aquatic Models

水生模型保存种质资源开发

• 批准号: 10457071
• 负责人: HARVEY D. BLACKBURN
• 金额: $ 6万
• 依托单位: LOUISIANA STATE UNIV AGRICULTURAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10457071
• 关键词: 3-Dimensional; 3D Print; Address; Animal Model; Animals; Back; Biological Testing; Biomedical Research; Communities; Community Developments; Coupling; Cryopreservation; Deposition; Development; Device or Instrument Development; Devices; Embryo; Engineering; Ensure; Equilibrium; Evaluation; Failure; Female; Fertilization; Fishes; Freezing; Genetic Research; Genotype; Glass; Goals; Guidelines; Health; Human; Individual; International; Investments; Label; Laboratories; Laboratory Research; Learning; Maintenance; Methods; Microfabrication; Modality; Modeling; Osmolalities; Output; Pathway interactions; Printing; Procedures; Process; Protocols documentation; Quality Control; Reporting; Reproducibility; Research; Resources; Risk; Safety; Sampling; Sperm Motility; Standardization; Techniques; Terminology; Testing; Time; Transgenic Organisms; Variant; Viscosity; Work; Zebrafish; aquarium; base; cell motility; cost; cost shifting; design; genetic resource; genome editing; human disease; improved; male; member; microdevice; mutant; neglect; novel strategies; open source; preservation; prevent; programs; repository; sample collection; sperm cell; success; wasting

Zebrafish (Danio rerio) is a powerful model in biomedical research and laboratories around the world have produced tens of thousands of mutant and transgenic lines. Maintaining these valuable genotypes as live fish is expensive, risky, and beyond the capacity of stock centers. As such, cryopreservation has become a necessity and most of these genetic resources are now maintained as samples of inconsistent quality frozen with rudimentary techniques. Quality control has not been practiced in any systematic way, reproducibility is poor, and protocols are not standardized. It is common to have problems and failures in fertilization resulting in lost lines that need to be recreated, causing facilities to waste considerable time and effort. This is largely due to the false notion that neglecting quality control saves time and money. However, rather than being reduced, these costs are shifted downstream through wasted storage space and reduced fertilization. This pervasive lack of quality control has placed the substantial investments in biomedical research at great risk. Therefore, our long-term goal is to provide inexpensive, universally available and systematic quality control leading to development of community-based standards for cryopreservation. This will enable reliable contributions from individual laboratories to large comprehensive repositories providing protection for genetic resources pivotal in biomedical research. To achieve this, we propose to improve reliability and efficiency by providing routine access to reproducibility and standardization through continued development of microfabricated (“laboratory on a chip”) and 3-dimensional (3-D) printed devices encompassing all process steps from sample collection through fertilization. The Specific Aims are to: 1) Develop and test microfabricated devices that can be used by research laboratories at low effort and cost to improve assessment and study of sperm concentration and motility with respect to the effects of these factors on reproducibility and the overall success of cryopreservation. 2) Develop and test 3-D printed devices that can be used to improve the reproducibility of the freezing process. These devices will address conventional cryopreservation and vitrification, and be useful for single samples and pooled batches. 3) Perform biological testing of these devices to refine design and function and improve the reproducibility of quality assessment to enable research laboratories to back up lines or submit them to stock centers and germplasm repositories. This will provide a community-based approach for protection of genetic resources through systematic incorporation of devices, guidelines and standards applicable across a full range of activity scopes and scales.

斑马鱼（Danio rerio）是生物医学研究和实验室的一个强大模型， 世界上已经产生了数以万计的突变体和转基因品系。维持这些 有价值的基因型如活鱼是昂贵的，有风险的，超出了库存中心的能力。作为 因此，冷冻保存已成为一种必需，这些遗传资源中的大多数现在 作为质量不一致的样品保存，用基本技术冷冻。质量 没有以任何系统的方式进行控制，再现性差，并且方案 不规范在施肥中出现问题和失败导致品系丢失是很常见的 需要重新创建，导致设施浪费大量时间和精力。这主要是 由于错误的观念，忽视质量控制节省时间和金钱。然而， 而不是减少，这些成本通过浪费的存储空间转移到下游， 减少施肥。这种普遍缺乏质量控制的情况使大量的 生物医学研究的投资风险很大。因此，我们的长期目标是提供 廉价、普遍可用和系统的质量控制， 基于社区的低温保存标准。这将使可靠的贡献， 从单个实验室到大型综合储存库， 生物医学研究中的关键资源。为了实现这一目标，我们建议提高可靠性， 通过持续提供可重复性和标准化的常规访问， 微加工（“芯片上的实验室”）和三维（3-D）打印设备的开发 包括从样品收集到受精的所有过程步骤。具体目标 是：1）开发和测试微加工设备，可用于研究实验室在 改善精子浓度和活力的评估和研究的工作量和成本较低， 考虑到这些因素对再现性和总体成功的影响， 冷冻保存2)开发和测试3D打印设备，可用于改善 冷冻过程的再现性。这些设备将解决传统的 冷冻保存和玻璃化，并可用于单一样品和合并批次。第三章 对这些器械进行生物学试验，以完善设计和功能，并改善 质量评估的可重复性，使研究实验室能够备份生产线或提交 将它们运送到储备中心和种质库。这将提供一个基于社区的 通过系统地采用装置保护遗传资源的方法， 适用于所有活动范围和规模的准则和标准。

项目成果

A microfluidic device for motility and osmolality analysis of zebrafish sperm.

• DOI: 10.1007/s10544-018-0308-2
• 发表时间: 2018-08-09
• 期刊: Biomedical microdevices
• 影响因子: 2.8
• 作者: Beckham J;Alam F;Omojola V;Scherr T;Guitreau A;Melvin A;Park DS;Choi JW;Tiersch TR;Todd Monroe W
• 通讯作者: Todd Monroe W

Workshop and panel discussion: high-throughput cryopreservation of germplasm as an exchange currency for genetic resources.

研讨会和小组讨论：种质高通量低温保存作为遗传资源的交换货币。

• DOI: 10.1016/j.cbpc.2011.06.008
• 发表时间: 2012
• 期刊: Comparative biochemistry and physiology. Toxicology & pharmacology : CBP
• 作者: Varga,ZoltánM;Tiersch,TerrenceR
• 通讯作者: Tiersch,TerrenceR

3-D printing provides a novel approach for standardization and reproducibility of freezing devices.

• DOI: 10.1016/j.cryobiol.2017.03.010
• 发表时间: 2017-06
• 期刊: Cryobiology
• 影响因子: 2.7
• 作者: Hu E;Childress W;Tiersch TR
• 通讯作者: Tiersch TR

Evaluation of cryoprotectant and cooling rate for sperm cryopreservation in the euryhaline fish medaka Oryzias latipes.

• DOI: 10.1016/j.cryobiol.2010.07.006
• 发表时间: 2010-10
• 期刊: Cryobiology
• 影响因子: 2.7
• 作者: Yang H;Norris M;Winn R;Tiersch TR
• 通讯作者: Tiersch TR

Determination of sperm concentration using flow cytometry with simultaneous analysis of sperm plasma membrane integrity in zebrafish Danio rerio.

使用流式细胞术测定斑马鱼斑马鱼精子浓度并同时分析精子质膜完整性。

• DOI: 10.1002/cyto.a.22796
• 发表时间: 2016
• 期刊: Cytometry. Part A : the journal of the International Society for Analytical Cytology
• 作者: Yang,Huiping;Daly,Jonathan;Tiersch,TerrenceR
• 通讯作者: Tiersch,TerrenceR