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标准慢速冷冻与无冷冻保护剂玻璃化冷冻后人精子的DNA完整性与活力

DNA integrity and motility of human spermatozoa after standard slow freezing versus cryoprotectant-free vitrification.

作者信息

Isachenko E, Isachenko V, Katkov I I, Rahimi G, Schöndorf T, Mallmann P, Dessole S, Nawroth F

机构信息

Department of Obstetrics and Gynecology, University of Cologne, Kerpener Str. 34, D-50931 Cologne, Germany.

出版信息

Hum Reprod. 2004 Apr;19(4):932-9. doi: 10.1093/humrep/deh194. Epub 2004 Mar 11.

Abstract

BACKGROUND

In contrast to the technique of conventional freezing, the vitrification of spermatozoa requires high cooling rates (720 000 degrees K/min), which could be damaging for spermatozoa. The aim of our study was to compare slowly frozen and vitrified spermatozoa in terms of their post-thaw DNA integrity and motility.

METHODS

Semen samples were prepared according to the routine swim-up technique and divided into aliquots for comparison of fresh, conventionally frozen and vitrified spermatozoa from the same ejaculate in the presence or absence of cryoprotectants. Spermatozoa motility and DNA integrity were determined.

RESULTS

The motility of spermatozoa conventionally (slowly) frozen with a cryoprotectant was similar to that recorded for spermatozoa vitrified in the absence of cryoprotectant (47 versus 52%). The DNA integrity was unaffected by the cryopreservation method or presence of cryoprotectants.

CONCLUSION

The vitrification of human spermatozoa in the absence of conventional cryoprotectants is indeed feasible. The DNA integrity of vitrified sperm is comparable with that shown by standard slow-frozen/thawed spermatozoa, yet the method is quick and simple and does not require special cryobiological equipment.

摘要

背景

与传统冷冻技术不同,精子玻璃化冷冻需要极高的降温速率(720000开尔文/分钟),这可能会对精子造成损害。我们研究的目的是比较缓慢冷冻和玻璃化冷冻精子解冻后的DNA完整性和活力。

方法

精液样本按照常规上游法制备,分成几份,用于比较同一射精样本中新鲜的、传统冷冻和玻璃化冷冻的精子,有无冷冻保护剂。测定精子活力和DNA完整性。

结果

使用冷冻保护剂进行传统(缓慢)冷冻的精子活力与未使用冷冻保护剂进行玻璃化冷冻的精子活力相似(分别为47%和52%)。DNA完整性不受冷冻保存方法或冷冻保护剂的影响。

结论

不使用传统冷冻保护剂对人类精子进行玻璃化冷冻确实可行。玻璃化冷冻精子的DNA完整性与标准缓慢冷冻/解冻精子相当,而且该方法快速简便,无需特殊的低温生物学设备。

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