CRYOPRESERVATION OF SMALL NUMBERS OF HUMAN SPERMATOZOA USING AGAROSE-GEL MICROCAPSULES
CRYOPRESERVATION OF SMALL NUMBERS OF HUMAN SPERMATOZOA USING AGAROSE-GEL MICROCAPSULES
复制标题
使用琼脂糖凝胶微胶囊冷冻保存少量人类精子
DOI：
10.1097/ju.0000000000000887.013
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
and Yoshitomo Kobori
中科院分区：
文献类型：
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作者：
Hiroshi Okada*;Toshiyuki Iwahata;Akiyoshi Osaka;and Yoshitomo Kobori
方法：精液样本收集自一所大学医院生殖医学中心男性不育门诊的16名患者。我们使用琼脂糖微胶囊冷冻保存了16例男性不育患者的极少量精子（10例精子浓度≥ 100万/mL; 6例精子浓度<100万/mL）。将6个精子注射入琼脂糖凝胶微囊中，并在液氮罐中冷冻保存7天。Crytop方法用于冷冻保存作为对照。解冻后，回收精子。结果：对精子浓度≥ 100万/mL的样本进行调查精子回收率：琼脂糖凝胶胶囊法为93%（279/300），Cryotop法为86%（257/300），两种方法的精子回收率有显著性差异（P< 0.005）。精子活率：琼脂糖胶囊法为73%（219/300），显著高于Cryotop法的51%（152/300）（P< 0.001）。精子存活率：琼脂糖胶囊法阳性率为86%（258/300），明显高于Cryotop法的65%（196/300）（P< 0.001）。精子回收时间（平均值±标准差）：琼脂糖凝胶胶囊法为3.56±1.5 min，明显短于Cryotop法的7.04±3.2 min（P< 0.001）。精子浓度小于100万/mL的样本的调查精子回收率：琼脂糖凝胶胶囊法为99%（179/180），显著高于Cryotop法的83%（150/180）（P< 0.001）。精子活率：琼脂糖胶囊法87%（157/180）显著高于Cryotop法69%（125/180）（P< 0.001）。精子存活率：琼脂糖胶囊法阳性率为86%（258/300），明显高于Cryotop法的65%（196/300）（P< 0.001）。精子回收时间（平均值±SD）：琼脂糖凝胶胶囊法为2.97±1.2 min，明显短于Cryotop法的8.20±2.9 min（P< 0.001）。上述结果表明，无论是精子浓度大于或等于100万/mL组，还是精子浓度小于100万/mL组，与Cryotop法相比，琼脂糖凝胶胶囊法保存的样本解冻后精子回收率、精子活率和精子存活率较高，而精子回收时间较短。本研究表明，与常规Cryotop方法相比，在冷冻保存精子计数低的样本时，使用琼脂糖凝胶胶囊方法可提高解冻后精子回收率、精子活动率和精子存活率，并缩短精子回收时间。因此，琼脂糖凝胶胶囊法可能是一种有前途的治疗男性不育症的方法。
METHODS:Semen samples were collected from 16 patients attending the male infertility clinic of the center for reproductive medicine at a university hospital. We used agarose micro capsules to cryopreserve extremely small numbers of sperm from 16 patients with male infertility (10 with sperm concentration≥ 1 million/mL; 6 with sperm concentration< 1 million/mL). Six spermatozoa were injected into agarose gel microcapsules and cryopreserved in a liquid nitrogen tank for 7 days. The Crytop method was used for cryopreservation as a control. After thawing, spermatozoa were recovered. Sperm recovery rates, motility and viability, and recovery time were compared.RESULTS:Investigation of samples with sperm concentration 1 million/mL or above Sperm recovery rate: 93%(279/300) for the agarose gel capsule method was significantly higher than the 86%(257/300) for the Cryotop method (P< 0.005). Sperm motility rate: 73%(219/300) for the agarose gel capsule method was significantly higher than the 51%(152/300) for the Cryotop method (P< 0.001). Sperm viability rate: 86%(258/300) for the agarose gel capsule method was significantly higher than the 65%(196/300) for the Cryotop method (P< 0.001). Sperm recovery time (mean±SD): 3.56±1.5 min for the agarose gel capsule method was significantly shorter than the 7.04±3.2 min for the Cryotop method (P< 0.001). Investigation of samples with sperm concentration less than 1 million/mL Sperm recovery rate: 99%(179/180) for the agarose gel capsule method was significantly higher than the 83%(150/180) for the Cryotop method (P< 0.001). Sperm motility rate: 87%(157/180) for the agarose gel capsule method was significantly higher than the 69%(125/180) for the Cryotop method (P< 0.001). Sperm viability rate: 86%(258/300) for the agarose gel capsule method was significantly higher than the 65%(196/300) for the Cryotop method (P< 0.001). Sperm recovery time (mean±SD): 2.97±1.2 min for the agarose gel capsule method was significantly shorter than the 8.20±2.9 min for the Cryotop method (P< 0.001). The results above showed that the post-thawing sperm recovery rate, sperm motility rate, and sperm viability rate were higher while the sperm recovery time was shorter in samples preserved using the agarose gel capsule method compared to samples preserved using the Cryotop method in both the group with sperm concentrations of 1 million/mL or above and the group with sperm concentrations of less than 1 million/mL.CONCLUSIONS:This study demonstrated that using the agarose gel capsule method increased post-thawing sperm recovery rate, sperm motility rate, and sperm viability rate, and reduced sperm recovery time compared with the conventional Cryotop method when cryopreserving samples with low sperm count. Thus, the agarose gel capsule method may be a promising treatment for male infertility.