Abstract:
The objectives of this study were to evaluate the effect of a short, cooled storage before cryopreservation on sperm progressive motility (PM) and compare the effect of different centrifugation methods on post-thaw PM of stored samples. Semen was diluted in chilling extender and aliquoted in 6 protocols: i) Standard centrifugation (SC) followed by freezing; ii) Single Layer Centrifugation (SLC) followed by freezing; iii) Storage for 8 h/5 °C before SC; iv) Storage for 8 h/5 °C before SLC; v) Storage for 8 h/15 °C before SC; and vi) Storage for 8 h/15 °C before SLC. PM was assessed before centrifugation, after centrifugation, and post-thawing. Stallions were classified as \"good freezers\" (GF) or \"bad freezers\" (BF). The PM in samples immediately frozen was greater than in the stored ones (71.98 ± 14.2, 52.91 ± 17.8, 53.93 ± 18.9 for no storage, 5 ºC storage and 15 ºC storage, respectively) (P˂ 0.0001). There was an effect of storage condition (p ˂ 0.0001), centrifugation method (p ˂ 0.0001), and freezability (P=0.0016), with an interaction between them (P= 0.0004), on PM after centrifugation. Post-thaw PM was greater in samples treated by SLC than in samples processed by SC, for all storage conditions (p ˂ 0.05). All BF stallions \'showed post-thaw PM ˂ 30 % when samples were previously stored. Storage at 5 ºC or 15º C for 8 h maintains an appropriate quality in GF stallions. Applying a sperm selection technique as SLC is suggested to improve post-thaw motility, allowing GF straws to be frozen after storage, although BF semen should be prepared by SLC immediately after collection.
摘要:
这项研究的目的是评估短期的影响,冷冻保存前冷却储存对精子进行性运动性(PM)的影响,并比较不同离心方法对储存样品解冻后PM的影响。将精液在冷冻增量剂中稀释并按6种方案等分:i)标准离心(SC),然后冷冻;ii)单层离心(SLC),然后冷冻;iii)在SC前储存8h/5°C;iv)在SLC前储存8h/5°C;v)在SC前储存8h/15°C;和vi)在SLC前储存8h/15°C。离心前评估PM,离心后,和解冻后。种马被归类为“好冰柜”(GF)或“坏冰柜”(BF)。立即冷冻样品的PM大于储存样品的PM(71.98±14.2,52.91±17.8,53.93±18.9,5ºC存储和15ºC存储,分别)(P<0.0001)。有储存条件的影响(p=0.0001),离心法(p=0.0001),和冻结性(P=0.0016),它们之间的相互作用(P=0.0004),离心后的PM。SLC处理的样品的解冻后PM高于SC处理的样品,适用于所有储存条件(p=0.05)。当样品先前储存时,所有BF种马显示解冻后PM为30%。在5ºC或15ºC下储存8小时可保持GF种马的适当质量。建议应用精子选择技术作为SLC来改善解冻后的运动性,允许GF吸管在储存后冷冻,虽然BF精液应在收集后立即由SLC制备。