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Effect of Dilution / Centrifugation on Yield and Motility of Cryopreserved Mouse Spermatozoa

机译：稀释/离心对冷冻保存的小鼠精子产量和运动性的影响

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Since mouse sperm cryopreservation has been achieved, progress has been marred by interstrain differences in post-thaw motility and fertility. A sensitivity difference to hyperosmosity among mouse strains could be one reason of this problem. The dilution of hyperosmotic cryoprotective agents was examined in BALB/c, C3H/HeN, and DBA/2N in order to improve post thaw motilities. Epididymal sperm were isolated and frozen in 18% raffinose containing buffer solution (～430mOsm/kg). The warmed samples were diluted with ×6 volume of BPS and centrifuged at 400×g 5min, 400×g 10min, 600×g 5min, 600×g 10min, and 800×g 10min. The number of sperm cells / clusters, and motilities were counted both in a supernatant and a sediment. After 400×g 5min centrifugation, 31,26, and 21 % of sperm were trapped in a supernatant in BALB/c, C3H/HeN, and DBA/2N, respectively, but the values were reduced to 22, 17, and 15 when centrifugation was accelerated to 600×g for 5min. The motilities were improved after centrifugation in 3 strains (but not significantly). We have reported the breakup of aggregated sperm by centrifugation significantly enhanced post thaw motility and fertility in cryopreserved C57BL strain (Cryobiology, 55 (2007), 340). Even the numbers of the clusters in each strain were reduced by centrifugation (significantly (p < 0.05) in BALB/c, C3H/HeN), absolute numbers of the clusters were extremely lower in each strains (only 1.2, 3.2, and 0.5 per 100 cells, which were less than a quarter of that in C57BL). So that the main cause of the improvement of motility could be a returning to isosmosity rather than the breakup of sperm clusters in these strains.
机译：由于已经实现了小鼠精子的冷冻保存，解冻后运动能力和受精能力之间的差异已经损害了进展。小鼠品系之间对高渗的敏感性差异可能是造成此问题的原因之一。在BALB / c，C3H / HeN和DBA / 2N中检查了高渗透性冷冻保护剂的稀释度，以提高解冻后的适应性。分离附睾精子，并在含18％棉子糖的缓冲液（〜430mOsm / kg）中冷冻。用×6体积的BPS稀释加热的样品，并在400×g 5min，400×g 10min，600×g 5min，600×g 10min和800×g 10min离心。计数上清液和沉淀物中的精子细胞/簇的数目和能动性。在400×g离心5分钟后，分别将31.26％和21％的精子截留在BALB / c，C3H / HeN和DBA / 2N的上清液中，但当精子的值降低到22、17和15时将离心加速至600×g 5分钟。在3个菌株中离心后，效用得到改善（但不显着）。我们已经报道了通过离心作用使聚集的精子破裂，这大大增强了冷冻保存的C57BL菌株的融解后运动能力和受精能力（Cryobiology，55（2007），340）。甚至通过离心减少了每个菌株中的簇数（在BALB / c，C3H / HeN中显着（p <0.05）），每个菌株中簇的绝对数也极低（每株仅1.2、3.2和0.5）。 100个单元，少于C57BL中的四分之一。因此，提高运动能力的主要原因可能是等渗的恢复，而不是这些菌株中精子簇的破裂。

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