Resources for Drosophila embryo cryopreservation at lab and stock center scale

实验室和库存中心规模的果蝇胚胎冷冻保存资源

• 批准号: 10569277
• 负责人: JOHN C BISCHOF
• 金额: $ 74.57万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10569277
• 关键词: Acceleration; Adoption; Adult; Animal Model; Benchmarking; Biomedical Research; Collaborations; Collection; Communities; Cost-Benefit Analysis; Costs and Benefits; Cryopreservation; Disasters; Drosophila genus; Drosophila melanogaster; Educational workshop; Embryo; Event; Feedback; Freezing; Future; Genetic; Genetic Diseases; Genetic Drift; Goals; Human Resources; Indiana; Individual; Larva; Maintenance; Malignant Neoplasms; Methods; Outcome; Outcomes Research; Pathway interactions; Population; Prevention; Procedures; Protocols documentation; Reagent; Reproducibility; Research; Research Personnel; Resource Sharing; Resources; Rewarming; Risk; Services; Shipping; Source; Standardization; Temperature; Testing; Time; Training; Work; cost; cost effective; cost estimate; cryobiology; cryogenics; embryo cryopreservation; experience; flexibility; fly; genetic resource; hatching; innovation; instrument; next generation sequencing; preservation; prevent; research and development; success; tool

ABSTRACT Drosophila melanogaster is one of the leading animal models for biomedical research, with researchers generating new lines of Drosophila every year for studies in numerous cancers, genetic disorders, and other maladies. Currently there are greater than 160,000 Drosophila stocks held at different stock centers around the world. As reliable and cost-effective approaches for long-term preservation of Drosophila stocks are lacking, individual labs and stock centers must maintain their lines as living populations, which is resource-intensive and puts the stocks at risk of loss. In 2021, our group developed an easily implemented and robust cryopreservation protocol for Drosophila melanogaster embryos which can be applied in any lab without the need for specialized instruments. This protocol is broadly applicable, and it has been successfully used to preserve 25 distinct strains from different sources. For most strains, >50% of the embryos hatch and >25% of the resulting larvae develop into fully functioning adults after cryopreservation and rewarming (normalized survival to control embryos), providing sufficient numbers of adults to revive the strain. To accelerate the cryopreservation and archival of critical Drosophila stocks, we will develop, standardize, and disseminate resources to individual labs and stock centers. Significant efforts will be devoted to developing reliable methods for cryogenic storage and shipping of Drosophila embryos as well as to study if any significant mutagenic changes occur due to cryopreservation. We will be working with multiple groups within the cryobiology and fly community to gather feedback on and refine our approach. With the collaboration of the Bloomington Drosophila Stock Center (BDSC), we will identify and prioritize the stocks that need cryopreservation to prevent their total loss in the event of a disaster or genetic drift. We will also demonstrate the efficacy of our cryopreservation protocol on a large scale with a selected group of stocks identified by BDSC. Finally, we will also estimate the costs for cryopreservation in comparison to the traditional method of Drosophila maintenance to inform future efforts to protect these precious genetic resources. This is a multi-PI project led by investigators with significant experience in Drosophila genetics and cryobiology.

抽象的 黑腹果蝇是生物医学研究的主要动物模型之一，研究人员 每年产生新的果蝇品系，用于研究多种癌症、遗传性疾病和其他疾病 疾病。目前，世界各地不同的库存中心持有超过 160,000 只果蝇库存。 世界。由于缺乏长期保存果蝇种群的可靠且具有成本效益的方法， 各个实验室和库存中心必须维持其作为活种群的线，这是资源密集型的，并且 使股票面临亏损的风险。 2021 年，我们的团队开发了一种易于实施且强大的冷冻保存方法 果蝇胚胎实验方案，可应用于任何实验室，无需专门的 仪器。该方案适用范围广泛，已成功用于保存 25 种不同的菌株 来自不同的来源。对于大多数菌株，>50% 的胚胎孵化，>25% 的幼虫发育 冷冻保存和复温后发育成功能齐全的成年人（正常化存活以控制胚胎）， 提供足够数量的成年人来恢复菌株。加快冷冻保存和归档 关键的果蝇种群，我们将开发、标准化资源并将其分发给各个实验室和种群 中心。将投入大量精力开发可靠的低温储存和运输方法 果蝇胚胎以及研究是否因冷冻保存而发生任何显着的诱变变化。我们 将与低温生物学和飞行界的多个小组合作，收集反馈并完善 我们的方法。在布卢明顿果蝇库存中心 (BDSC) 的合作下，我们将识别并 优先考虑需要冷冻保存的种群，以防止在发生灾难或遗传事件时全部损失 漂移。我们还将用选定的方法大规模展示我们的冷冻保存方案的有效性 BDSC 确定的股票组。最后，我们还会比较一下冷冻保存的成本 果蝇的传统维护方法，为未来保护这些珍贵遗传的努力提供信息 资源。这是一个多 PI 项目，由在果蝇遗传学和遗传学领域拥有丰富经验的研究人员领导。 低温生物学。