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， 每年产生新的果蝇品系，用于研究许多癌症、遗传疾病和其他疾病。 疾病目前有超过160，000果蝇股票持有在不同的股票中心周围 世界由于缺乏长期保存果蝇种群的可靠和具有成本效益的方法， 各个实验室和储备中心必须将其生产线作为活种群来维持，这是资源密集型的， 使股票面临亏损的风险。在2021年，我们的团队开发了一种易于实施和强大的冷冻保存方法， 黑腹果蝇胚胎的实验方案，可以在任何实验室应用，而不需要专门的 仪器.该方法具有广泛的适用性，已成功地用于保存25种不同的菌株 从不同的来源。对于大多数菌株，>50%的胚胎孵化，>25%的幼虫发育 在冷冻保存和复温后发育成功能完全的成年人（对照胚胎的标准化存活率）， 提供足够数量的成虫来恢复菌株。为了加速冷冻保存和存档 关键的果蝇种群，我们将开发，标准化和传播资源，以个别实验室和股票 中心.将作出重大努力，开发可靠的低温储存和运输方法， 果蝇胚胎以及研究是否由于冷冻保存而发生任何显著的致突变变化。我们 将与低温生物学和苍蝇社区内的多个小组合作，收集反馈意见， 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，以防止它们在发生灾难或遗传病时完全丧失。 漂移我们还将证明我们的低温保存方案的有效性，在大规模与选定的 BDSC确定的种群。最后，我们也将估计成本为冷冻保存的比较 果蝇的传统维护方法，以告知未来的努力，以保护这些宝贵的遗传 资源这是一个多PI项目，由在果蝇遗传学方面具有丰富经验的研究人员领导， 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.