Allen C H, Almquist J O
J Anim Sci. 1981 Dec;53(6):1432-9. doi: 10.2527/jas1982.5361432x.
Twelve ejaculates were extended both in skimmilk and Tris-yolk, packaged in .5-ml French straws and frozen vertically in bulk (10,000 straws/freeze) in a programmed mechanical freezer (Linde CRFC-3). Cooling rates and post-thaw spermatozoal survival in the upper and lower thirds of straws at the following six chamber positions were compared: corner and middle within-goblet positions at center, intermediate and outer chamber locations. Cooling rates generally were faster in the upper third of straws than in the lower third and at the corner rather than the middle within-goblet positions. For combined postthaw incubation periods of 0, 3 and 6 hr at 37 C, motility (photographic and visual) and acrosomal retention (fixed and unfixed samples) were both higher (P greater than .01) for sperm in each extender in the upper third of straws than in the lower third. Spermatozoal motility and acrosomal retention generally were higher (P greater than .05) in each extender at the center rather than at the outer chamber location. Among within-goblet positions, post-thaw survival of sperm was higher (P greater than .05) at the corner than at the middle positions for Tris-yolk but not skimmilk. For comparison semen was frozen in static N vapor in straws held singly (258-straw load) on horizontal racks. For combined incubation periods, post-thaw survival of sperm did not differ (P less than .05) between bulk and static vapor- systems. There also was no difference when each of the six chamber positions was compared individually with static vapor. In conclusion, 33% more straws of semen can be frozen in a mechanical freezer than previously reported, with post-thaw spermatozoal survival comparable to that of sperm in straws frozen conventionally in static vapor.
采集了12份精液样本,分别用脱脂乳和Tris-卵黄进行稀释,装入0.5毫升的法式细管中,并在程序控制的机械冷冻机(林德CRFC - 3型)中垂直批量冷冻(每次冷冻10,000根细管)。比较了在以下六个腔室位置的细管上三分之一和下三分之一处的冷却速率以及解冻后精子的存活率:中心杯状位置的角落和中间、中间腔室位置和外侧腔室位置。细管上三分之一处的冷却速率通常比下三分之一处快,且在杯状位置的角落而非中间位置更快。在37℃下解冻后分别孵育0、3和6小时,细管上三分之一处的每种稀释剂中的精子活力(通过摄影和肉眼观察)和顶体保留率(固定和未固定样本)均高于下三分之一处(P>0.01)。在每个稀释剂中,中心位置的精子活力和顶体保留率通常高于外侧腔室位置(P>0.05)。在杯状位置内,Tris-卵黄稀释的精子解冻后在角落位置的存活率高于中间位置(P>0.05),但脱脂乳稀释的情况并非如此。作为对照,精液在水平架上单独放置的细管(258根细管的装载量)中于静态氮蒸气中冷冻。对于合并的孵育期,批量冷冻和静态蒸气系统解冻后精子的存活率没有差异(P<0.05)。将六个腔室位置中的每一个与静态蒸气单独比较时也没有差异。总之,在机械冷冻机中冷冻的精液细管数量比之前报道的多33%,解冻后精子的存活率与传统在静态蒸气中冷冻的细管中的精子相当。