National Research Council (IBCN), CNR-Campus International Development (EMMA-INFRAFRONTIER-IMPC), Monterotondo Scalo, Rome, Italy.
Mary Lyon Centre, MRC Harwell, Harwell Science and Innovation Campus, Oxfordshire, OX11 0RD, United Kingdom.
Theriogenology. 2017 Jul 1;96:49-57. doi: 10.1016/j.theriogenology.2017.04.003. Epub 2017 Apr 3.
Disseminating mouse stocks as frozen materials offers both ethical and logistical advantages over live animal shipment, minimizing the welfare issues and avoiding some of the complex custom regulations that are associated with live animal transportation. Embryo freezing in liquid nitrogen (LN) at -196 °C has traditionally been the method of choice for archiving mouse lines. However, spermatozoa freezing is emerging as a more convenient alternative due to the application of innovative cryopreservation and recovery protocols. In addition, frozen spermatozoa are less sensitive to post-freezing temperature fluctuations. Here we demonstrated that spermatozoa frozen using standard laboratory protocols can be safely stored in dry ice (-79 °C) for at least seven days. The protocol we report here is robust and has been validated in a multi-centric study involving mouse spermatozoa samples exchanged between five European Mouse Mutant Archive (EMMA) nodes. Furthermore, following shipment on dry ice the spermatozoa can be returned to LN for long term storage without any noticeable detrimental effect. This protocol permits frozen spermatozoa to be shared and shipped in dry ice between biorepositories, networks and scientific institutions at low cost, using common courier companies, while avoiding the complexities, risks and hazards associated with using a traditional LN dry-shipper.
将小鼠品系以冷冻材料的形式传播，具有伦理和物流方面的优势，优于活体动物运输，可以最大限度地减少福利问题，并避免与活体动物运输相关的一些复杂的海关规定。在-196°C 的液氮（LN）中冷冻胚胎一直是保存小鼠品系的首选方法。然而，由于创新性的冷冻保存和复苏方案的应用，精子冷冻正在成为一种更方便的替代方法。此外，冷冻精子对冷冻后温度波动的敏感性较低。在这里，我们证明了使用标准实验室方案冷冻的精子可以安全地在干冰（-79°C）中储存至少七天。我们在这里报告的方案是稳健的，并已在涉及五个欧洲小鼠突变体档案（EMMA）节点之间交换的小鼠精子样本的多中心研究中得到验证。此外，在干冰运输后，精子可以返回 LN 进行长期储存，而不会产生任何明显的不利影响。该方案允许使用常见的快递公司，以低成本在生物库、网络和科学机构之间以干冰形式共享和运输冷冻精子，同时避免使用传统 LN 干贮箱相关的复杂性、风险和危害。
