Iryna Kuznyetsova, Sergey I Moskovtsev, Stephanie Ng, Bill Yee, Ayub G M Lulat, Valeriy Kuznyetsov, Clifford L Librach
{"title":"Permeable cryoprotectants-free vitrification of human TESE, PESA and OAT spermatozoa: clinical outcomes.","authors":"Iryna Kuznyetsova, Sergey I Moskovtsev, Stephanie Ng, Bill Yee, Ayub G M Lulat, Valeriy Kuznyetsov, Clifford L Librach","doi":"10.1080/19396368.2025.2466687","DOIUrl":null,"url":null,"abstract":"<p><p>Cryopreservation of testicular and epididymal spermatozoa is more challenging in comparison to ejaculated spermatozoa due to lower sperm concentration and motility, and higher sperm sensitivity to cryoprotectants. Sperm vitrification without the use of potentially toxic permeable cryoprotectants is an attractive freezing alternative for testicular and epididymal spermatozoa, as well as oligoasthenoteratozoospermia (OAT) samples. Our study is a retrospective analysis of outcomes in IVF cycles involving a total of 70 testicular, 77 epididymal and 69 ejaculated OAT samples vitrified in a closed double-straw device using mHTF medium supplemented with protein and sucrose, without any permeable cryoprotectant. In total, 71 frozen samples were used for intracytoplasmic sperm injection (ICSI). Results were compared to fresh samples (26 testicular, 53 epididymal and 63 ejaculated OAT samples) that served as controls. Elective single frozen embryo transfers of euploid or unknown-ploidy blastocysts were performed. While sperm motility is expected to diminish following slow sperm freezing and thawing, our data demonstrated that vitrification of testicular, epididymal and OAT samples had a mean motility rate comparable to fresh samples. No statistically significant differences (<i>p</i> > 0.05) were observed between vitrified versus fresh TESE in fertilization (64.1% vs. 59.5%), blastocyst development (54.9% vs. 56.7%), blastocyst euploidy (36.4% vs. 33.3%), clinical pregnancy (47.8% vs. 36.4%) and live birth rates (43.5% vs. 24.2%). Similarly, vitrified versus fresh PESA showed no statistically significant differences (<i>p</i> > 0.05) in the analyzed results respectively: (69.4% vs.74.9%; 62.6% vs. 59.7%; 40.5% vs. 48.1%; 36.0% vs.37.7%; and 32.0% vs. 27.5%). For vitrified OAT samples, there was a significant difference in blastocyst development and euploidy rates when compared to the control group. Our results demonstrate that human testicular, epididymal spermatozoa and samples with OAT can be successfully vitrified in small volumes in a closed system without using any permeable cryoprotectants, allowing utilization of this technique in clinical settings.</p>","PeriodicalId":22184,"journal":{"name":"Systems Biology in Reproductive Medicine","volume":"71 1","pages":"54-60"},"PeriodicalIF":2.1000,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Systems Biology in Reproductive Medicine","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1080/19396368.2025.2466687","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/2/21 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"ANDROLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Cryopreservation of testicular and epididymal spermatozoa is more challenging in comparison to ejaculated spermatozoa due to lower sperm concentration and motility, and higher sperm sensitivity to cryoprotectants. Sperm vitrification without the use of potentially toxic permeable cryoprotectants is an attractive freezing alternative for testicular and epididymal spermatozoa, as well as oligoasthenoteratozoospermia (OAT) samples. Our study is a retrospective analysis of outcomes in IVF cycles involving a total of 70 testicular, 77 epididymal and 69 ejaculated OAT samples vitrified in a closed double-straw device using mHTF medium supplemented with protein and sucrose, without any permeable cryoprotectant. In total, 71 frozen samples were used for intracytoplasmic sperm injection (ICSI). Results were compared to fresh samples (26 testicular, 53 epididymal and 63 ejaculated OAT samples) that served as controls. Elective single frozen embryo transfers of euploid or unknown-ploidy blastocysts were performed. While sperm motility is expected to diminish following slow sperm freezing and thawing, our data demonstrated that vitrification of testicular, epididymal and OAT samples had a mean motility rate comparable to fresh samples. No statistically significant differences (p > 0.05) were observed between vitrified versus fresh TESE in fertilization (64.1% vs. 59.5%), blastocyst development (54.9% vs. 56.7%), blastocyst euploidy (36.4% vs. 33.3%), clinical pregnancy (47.8% vs. 36.4%) and live birth rates (43.5% vs. 24.2%). Similarly, vitrified versus fresh PESA showed no statistically significant differences (p > 0.05) in the analyzed results respectively: (69.4% vs.74.9%; 62.6% vs. 59.7%; 40.5% vs. 48.1%; 36.0% vs.37.7%; and 32.0% vs. 27.5%). For vitrified OAT samples, there was a significant difference in blastocyst development and euploidy rates when compared to the control group. Our results demonstrate that human testicular, epididymal spermatozoa and samples with OAT can be successfully vitrified in small volumes in a closed system without using any permeable cryoprotectants, allowing utilization of this technique in clinical settings.
与射精精子相比，睾丸和附睾精子的冷冻保存更具挑战性，因为精子浓度和活力较低，精子对冷冻保护剂的敏感性较高。不使用具有潜在毒性的可渗透冷冻保护剂的精子玻璃化是睾丸和附睾精子以及少弱异卵精子症（OAT）样本的一种有吸引力的冷冻选择。我们的研究是对IVF周期的结果进行回顾性分析，共涉及70例睾丸，77例附睾和69例射精OAT样本，使用添加蛋白质和蔗糖的mHTF培养基，在没有任何渗透性冷冻保护剂的情况下，在封闭的双吸管装置中玻璃化。共71例冷冻标本用于胞浆内单精子注射（ICSI）。结果与作为对照的新鲜样本（26例睾丸，53例附睾和63例射精OAT样本）进行了比较。选择单冷冻胚胎移植整倍体或未知倍体囊胚。虽然精子活力在缓慢的精子冷冻和解冻后会下降，但我们的数据表明，玻璃化的睾丸、附睾和OAT样本的平均活力率与新鲜样本相当。玻璃化TESE与新鲜TESE在受精率（64.1% vs. 59.5%）、囊胚发育（54.9% vs. 56.7%）、囊胚整倍性（36.4% vs. 33.3%）、临床妊娠率（47.8% vs. 36.4%）和活产率（43.5% vs. 24.2%）方面无统计学差异（p < 0.05）。同样，玻璃化的PESA与新鲜的PESA在分析结果中差异无统计学意义（p > 0.05）： 69.4% vs.74.9%；62.6% vs. 59.7%；40.5%对48.1%；vs.37.7% 36.0%;32.0% vs. 27.5%)。对于玻璃化的OAT样品，与对照组相比，囊胚发育和整倍体率有显着差异。我们的研究结果表明，人类睾丸，附睾精子和OAT样品可以在封闭系统中成功地小体积玻璃化，而无需使用任何渗透性冷冻保护剂，从而允许在临床环境中使用该技术。
期刊介绍：
Systems Biology in Reproductive Medicine, SBiRM, publishes Research Articles, Communications, Applications Notes that include protocols a Clinical Corner that includes case reports, Review Articles and Hypotheses and Letters to the Editor on human and animal reproduction. The journal will highlight the use of systems approaches including genomic, cellular, proteomic, metabolomic, bioinformatic, molecular, and biochemical, to address fundamental questions in reproductive biology, reproductive medicine, and translational research. The journal publishes research involving human and animal gametes, stem cells, developmental biology and toxicology, and clinical care in reproductive medicine. Specific areas of interest to the journal include: male factor infertility and germ cell biology, reproductive technologies (gamete micro-manipulation and cryopreservation, in vitro fertilization/embryo transfer (IVF/ET) and contraception. Research that is directed towards developing new or enhanced technologies for clinical medicine or scientific research in reproduction is of significant interest to the journal.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
