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人类精子的冷冻保存:两种冷冻保存方法与三种冷冻保护剂的比较

Cryopreservation of human spermatozoa: comparison of two cryopreservation methods and three cryoprotectants.

作者信息

Nallella Kiran P, Sharma Rakesh K, Allamaneni Shyam S R, Aziz Nabil, Agarwal Ashok

机构信息

Center for Advanced Research in Human Reproduction, Infertility and Sexual Function, Glickman Urological Institute and Department of Obstetrics-Gynecology, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.

出版信息

Fertil Steril. 2004 Oct;82(4):913-8. doi: 10.1016/j.fertnstert.2004.02.126.

Abstract

OBJECTIVE

To evaluate the ability of two cryopreservation methods and three cryoprotectants to preserve sperm quality.

DESIGN

A prospective clinical study.

SETTING

Male infertility clinic at a tertiary healthcare center.

PATIENT(S): Twenty infertile men and 10 healthy donors.

INTERVENTION(S): In the first experiment, semen was cryopreserved by either the Irvine Scientific method (IS) or the Cleveland Clinic Foundation (CCF) method. In the second experiment, semen was cryopreserved by the IS method and one of three cryoprotectants: TES and Tris yolk buffer, Sperm Freezing Medium, or Enhance Sperm Freeze.

MAIN OUTCOME MEASURE(S): Postthaw sperm motility, cryosurvival, and kinematics.

RESULT(S): Percentages of postthaw sperm motility and cryosurvival were higher in the IS cryopreservation method compared with in the CCF method (15.94 +/- 9.19 vs. 12.07 +/- 7.31 and 47.42 +/- 17.44 vs. 35.76 +/- 17.56). However, the CCF method resulted in significantly better sperm kinematics. Postthaw motility in the donors and patients was highest in the samples frozen in TES and Tris yolk buffer medium.

CONCLUSION(S): The IS method was associated with more flash freezing compared with the CCF method and resulted in better preservation of sperm motility and a higher cryosurvival rate. TES and Tris yolk buffer was most effective at protecting sperm from the negative effects of the cryopreservation process. This may be due to the presence of egg yolk along with glycerol.

摘要

目的

评估两种冷冻保存方法和三种冷冻保护剂对精子质量的保存能力。

设计

前瞻性临床研究。

地点

三级医疗中心的男性不育诊所。

患者

20名不育男性和10名健康供者。

干预措施

在第一个实验中,精液采用欧文科学方法(IS)或克利夫兰诊所基金会(CCF)方法进行冷冻保存。在第二个实验中,精液采用IS方法和三种冷冻保护剂之一进行冷冻保存:TES和Tris蛋黄缓冲液、精子冷冻培养基或增强精子冷冻剂。

主要观察指标

解冻后精子活力、冷冻存活率和运动学参数。

结果

与CCF方法相比,IS冷冻保存方法解冻后精子活力百分比和冷冻存活率更高(分别为15.94±9.19对12.07±7.31以及47.42±17.44对35.76±17.56)。然而,CCF方法的精子运动学参数明显更好。在供者和患者中,解冻后活力在TES和Tris蛋黄缓冲液培养基中冷冻的样本中最高。

结论

与CCF方法相比,IS方法的快速冷冻更多,能更好地保存精子活力并提高冷冻存活率。TES和Tris蛋黄缓冲液在保护精子免受冷冻保存过程的负面影响方面最有效。这可能是由于蛋黄与甘油同时存在的缘故。

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