Resources for Drosophila embryo cryopreservation at lab and stock center scale

摘要 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、遗传疾病和其他 疾病。目前，有超过16万只果蝇股票存放在世界各地的不同股票中心 世界。由于缺乏可靠和具有成本效益的方法来长期保存果蝇种群， 单个实验室和库存中心必须保持其作为活人口的生产线，这是资源密集型和 使股票面临损失的风险。2021年，我们的团队开发出了一种易于实施且功能强大的冷冻保存方法 适用于任何实验室的黑腹果蝇胚胎方案，不需要专门的 乐器。该协议具有广泛的适用性，并已成功地用于保存25个不同的菌株 来自不同的来源。对于大多数品系，50%的胚胎孵化，25%的幼虫发育。 在冷冻保存和复温(正常存活以控制胚胎)后变成完全功能的成体， 提供了足够数量的成虫来重振这种菌株。为加快超低温保存和归档 关键果蝇种群，我们将开发、标准化并将资源分发给各个实验室和种群 中锋。将致力于开发可靠的低温储存和运输方法 此外，我们还将继续研究果蝇胚胎，并研究冷冻保存是否会发生任何重大的突变变化。我们 将与低温生物学和飞行社区内的多个小组合作，收集反馈并改进 我们的方法。在布鲁明顿果蝇种群中心(BDSC)的合作下，我们将识别和 对需要超低温保存的库存进行优先排序，以防止它们在发生灾难或基因突变时完全丢失 漂移。我们还将在大范围内通过选定的 由BDSC确定的股票组。最后，我们还将对冷冻保存的成本进行比较 以传统的方法维护果蝇，通知未来努力保护这些珍贵的基因 资源。这是一个多PI项目，由在果蝇遗传学和 低温生物学。

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.