Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: Effecy on motility, DNA integrity, and fertilization ability
Cryoprotectant-free cryopreservation of human spermatozoa by vitrification and freezing in vapor: Effecy on motility, DNA integrity, and fertilization ability
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DOI：
10.1095/biolreprod.104.028811
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发表时间：
2004-10-01
影响因子：
3.6
通讯作者：
van der Ven, H
中科院分区：
文献类型：
--
作者：
Isachenko, V;Isachenko, E;van der Ven, H
人类精子可以在不使用冷冻保护剂的情况下，以非常高的冷却速率（高达7.2 × 10（5）℃/min）进行玻璃化冷冻，从而成功地进行冷冻保存。这是通过将装有精子悬浮液的铜冷冻环直接投入液氮中来实现的。储存后，玻璃化的精子通过在搅拌的温暖介质中融化而立即解冻。本研究的目的是比较使用这种快速玻璃化冷冻方法冷冻保存的精子的质量与在浸入液氮之前将装载的冷冻环预暴露于液氮蒸气（-160 ℃）中以150- 250 ℃/min的速度冷却的精子的质量。这两种冷却模式导致的运动性，受精能力，和DNA完整性的温暖的精子方面的可比结果。在这两种情况下，在温暖的介质中融化的瞬间解冻是成功冷冻保存的必要条件。我们的研究结果表明，冷冻保护剂的冷冻保存精子的最佳方案不需要限制在液氮中储存前非常快的冷却，广泛的冷却速率是可以接受的。在此，我们讨论了这一发现的影响，在光的物理学的外和细胞内玻璃化。
Human spermatozoa can be successfully cryopreserved avoiding the use of cryoprotectants through vitrification at very high cooling rates (up to 7.2 x 10(5) degreesC/min). This is achieved by directly plunging a copper cryoloop loaded with a sperm suspension into liquid nitrogen. After storage, vitrified spermatozoa are instantly thawed by melting in an agitated, warm medium. The goal of the present study was to compare the quality of spermatozoa cryopreserved using this rapid vitrification method with that of spermatozoa cooled relatively slowly by preexposure of the loaded cryoloop to liquid nitrogen vapor (-160degreesC) with speed in the range 150-250degreesC/min) before immersion into liquid nitrogen. Both cooling modes led to comparable results in terms of the motility, fertilization ability, and DNA integrity of the warmed spermatozoa. In both cases, instant thawing by melting in a warm medium was essential for successful cryopreservation. Our findings suggest that optimal regimes for the cryoprotectant-free cryopreservation of spermatozoa need not be restricted to very fast cooling before storage in liquid nitrogen, a wide range of cooling rates being acceptable. Herein, we discuss the implications of this finding in the light of the physics of extra- and intracellular vitrification.