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基于磷脂酰胆碱和L-乙酰肉碱的冷冻培养基可以替代蛋黄并保留人类精子功能。

Phosphatidylcholine and L-acetyl-carnitine-based freezing medium can replace egg yolk and preserves human sperm function.

作者信息

Sicchieri Fernanda, Silva Aline Bomfim, Santana Viviane Paiva, Vasconcelos Maria Aparecida Carneiro, Ferriani Rui Alberto, Vireque Alessandra Aparecida, Dos Reis Rosana Maria

机构信息

Department of Gynecology and Obstetrics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.

National Institutes of Hormones and Woman's Health, CNPq, Brazil.

出版信息

Transl Androl Urol. 2021 Jan;10(1):397-407. doi: 10.21037/tau-20-1004.

Abstract

BACKGROUND

Conventional cryopreservation methods induce chemical and mechanical damage to the sperm membranes. The cryoprotectant potential of phospholipids of vegetal origin as soybean lecithin has been investigated as a substitute for egg yolk in diluents used for the cryopreservation of human spermatozoa. Therefore, the objective of this study was comparing the efficacy of a synthetic cryoprotectant supplemented with L-α-phosphatidylcholine (PC) and L-acetyl-carnitine (ANTIOX-PC) and the standard egg-based TEST-yolk buffer (TYB) in preserving sperm motility and chromatin quality in cryopreserved semen samples.

METHODS

Prospective experimental study in which semen samples from 63 men with normal sperm motility and 58 men with low sperm motility were included and analyzed both before and after cryopreservation using ANTIOX-PC or TYB freezing media. Sperm quality was evaluated by routine semen analysis and DNA fragmentation index using the Terminal deoxynucleotidyl transferase dUTP nick end labeling assay.

RESULTS

Differences in the post-thaw progressive motility and DNA fragmentation index were not detected between TYB and ANTIOX-PC cryoprotectants in both normal and low sperm motility groups (P>0.05). However, ANTIOX-PC medium retained higher non-progressive motility and lower percentage of immotile sperm when compared to TYB medium, resulting in a greater total motile sperm count (P<0.05), regardless baseline values of motility characteristic of the normospermic or asthenozoospermic samples.

CONCLUSIONS

ANTIOX-PC medium was effective to protect human sperm during a freeze-thaw cycle compared to the TYB medium. A clinically relevant advantage in better preserving kinetic parameters as higher total motility and lower immotile post-thawed sperm from ANTIOX-PC, in normal and low motility semen samples, demonstrated the positive impact of phospholipid and antioxidant treatment on sperm cryotolerance with high potential for egg yolk lipids replacement and biosafety.

摘要

背景

传统的冷冻保存方法会对精子膜造成化学和机械损伤。植物来源的磷脂如大豆卵磷脂作为冷冻保护剂的潜力已被研究,可替代用于人类精子冷冻保存的稀释剂中的蛋黄。因此,本研究的目的是比较添加L-α-磷脂酰胆碱(PC)和L-乙酰肉碱(ANTIOX-PC)的合成冷冻保护剂与标准的基于蛋黄的TEST-蛋黄缓冲液(TYB)在保存冷冻精液样本中精子活力和染色质质量方面的效果。

方法

一项前瞻性实验研究,纳入了63名精子活力正常的男性和58名精子活力低的男性的精液样本,使用ANTIOX-PC或TYB冷冻介质在冷冻保存前后进行分析。通过常规精液分析和使用末端脱氧核苷酸转移酶dUTP缺口末端标记法的DNA碎片化指数评估精子质量。

结果

在正常和低精子活力组中,TYB和ANTIOX-PC冷冻保护剂之间解冻后渐进性活力和DNA碎片化指数均未检测到差异(P>0.05)。然而,与TYB介质相比,ANTIOX-PC介质保留了更高的非渐进性活力和更低的不活动精子百分比,导致总活动精子数更高(P<0.05),无论正常精子或弱精子症样本活力特征的基线值如何。

结论

与TYB介质相比,ANTIOX-PC介质在冻融循环中对保护人类精子有效。在正常和低活力精液样本中,ANTIOX-PC在更好地保存动力学参数(如更高的总活力和更低的解冻后不活动精子)方面具有临床相关优势,证明了磷脂和抗氧化剂处理对精子冷冻耐受性的积极影响,具有很高的蛋黄脂质替代潜力和生物安全性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f33/7844480/d27f087bda19/tau-10-01-397-f1.jpg

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