Sieme Harald, Oldenhof Harriëtte
Clinic for Horses-Unit for Reproductive Medicine, University of Veterinary Medicine Hannover, Bünteweg 15, D-30559, Hannover, Germany,
Methods Mol Biol. 2015;1257:277-87. doi: 10.1007/978-1-4939-2193-5_10.
In modern livestock breeding, cryopreserved semen is routinely used for artificial insemination. Sperm cryopreservation secures future reproduction, and insemination doses can be easily shipped. Processing of semen for cryopreservation can be done with minimal efforts and relatively low costs. In this chapter we describe the entire cryopreservation process for stallion and bull sperm including dilution of sperm in primary and freezing extender, cooling and packaging in straws, freezing in liquid nitrogen vapor, cryogenic storage, and thawing. Special emphasis is given on preparation of commonly used primary and freezing extenders (skim milk extender "INRA-82", TRIS-egg yolk extender "TEY") used in a two-step dilution approach. Furthermore the different cooling rates needed in different temperature regimes during the freezing process are being described. Cryopreservation procedures are described in case of using both specialized automated equipment and simple equipment.
在现代家畜育种中,冷冻精液常规用于人工授精。精子冷冻保存确保了未来的繁殖,并且授精剂量易于运输。精液冷冻保存的处理可以轻松完成且成本相对较低。在本章中,我们描述了种马和公牛精子的整个冷冻保存过程,包括精子在初级和冷冻稀释液中的稀释、在细管中冷却和包装、在液氮蒸气中冷冻、低温储存和解冻。特别强调了用于两步稀释法的常用初级和冷冻稀释液(脱脂奶稀释液“INRA - 82”、TRIS - 蛋黄稀释液“TEY”)的制备。此外,还描述了冷冻过程中不同温度区间所需的不同冷却速率。介绍了使用专门自动化设备和简单设备时的冷冻保存程序。