Intracytoplasmic sperm injection outcomes with cryopreserved testicular sperm aspiration samples

Intracytoplasmic sperm injection (ICSI) may be performed with testicular frozen–thawed spermatozoa in patients with nonobstructive azoospermia (NOA). Sperm retrieval can be performed in advance of oocyte aspiration, as it may avoid the possibility of no recovery of spermatozoa on the day of oocyte pickup. There are few studies available in the literature concerning the use of frozen–thawed spermatozoa obtained from testicular sperm aspiration (TESA). To evaluate the effects and the outcomes of ICSI with frozen–thawed spermatozoa obtained by TESA, we performed a retrospective analysis of 43 ICSI cycles using frozen–thawed TESA. We obtained acceptable results with a fertilisation rate of 67.9%, an implantation rate (IR) of 17.1%, and clinical and ongoing pregnancy rates of 41.9% and 37.2% respectively. The results of this study suggest that performing ICSI using cryopreserved frozen–thawed testicular spermatozoa with TESA as a first option is a viable, safe, economic and effective method for patients with NOA.     

Outcome of microsurgical testicular sperm extraction in familial idiopathic nonobstructive azoospermia

The aim of the study was to evaluate the sperm retrieval rate by microsurgical testicular sperm extraction (TESE) in familial idiopathic nonobstructive azoospermia (NOA). One hundred and nineteen patients with idiopathic NOA who underwent microsurgical TESE over the past 5 years were included. Patients were then divided into two groups; Group ‘A’ with familial idiopathic NOA (11 families with two brothers in each family, 22 patients) and Group ‘B’ with nonfamilial idiopathic NOA (97 patients). Clinical data as well as data of microsurgical TESE were recorded. In Group ‘A’, the sperm retrieval rate was 9.1% (2/22 patients) compared to 45.4% in Group ‘B’ (44/97 patients) (P ≤ 0.05). The two patients in Group ‘A’ with successful sperm retrieval belonged to one family. The histopathological diagnosis was the same in the brothers in each family. It can be concluded that the testicular sperm retrieval rate in familial idiopathic NOA is significantly lower than in nonfamilial idiopathic NOA.     

Paternal age as an independent factor does not affect embryo quality and pregnancy outcomes of testicular sperm extraction‐intracytoplasmic sperm injection in azoospermia

This study was performed to evaluate the independent influence of paternal age affecting embryo development and pregnancy using testicular sperm extraction (TESE)‐intracytoplasmic sperm injection (ICSI) in obstructive azoospermia (OA) and nonobstructive azoospermia (NOA). Paternal patients were divided into the following groups: ≤30 years, 31–35 years, 36–40 years, 41–45 years and ≥46 years. There were no differences in the rates of fertilisation or embryo quality according to paternal and maternal age. However, clinical pregnancy and implantation rates were significantly lower between those ≥46 years of paternal age compared with other age groups. Fertilisation rate was higher in the OA than the NOA, while embryo quality, pregnancy and delivery results were similar. Clinical pregnancy and implantation rates were significantly lower for patients ≥46 years of paternal age compared with younger age groups. In conclusion, fertilisation using TESE in azoospermia was not affected by the independent influence of paternal age; however, as maternal age increased concomitantly with paternal age, rates of pregnancy and delivery differed between those with paternal age <41 years and ≥46 years. Therefore, paternal age ≥46 years old should be considered when applying TESE‐ICSI in cases of azoospermia, and patients should be advised of the associated low pregnancy rates.     

Parameters predicting sperm retrieval rates during microscopic testicular sperm extraction in nonobstructive azoospermia

The aim of this study was to investigate the predictive value of the parameters that might have an effect on the success of microscopic testicular sperm extraction (micro‐TESE) in infertile patients with nonobstructive azoospermia (NOA). Between 2003 and 2014, 860 patients with NOA were retrospectively analysed. The effect of age, infertility duration, history of varicocelectomy, herniorrhaphy or orchiopexy, presence of solitary testis, tobacco use, previous testicular biopsy results, history of orchitis, usage of human chorionic gonadotropin in the past three months, presence of undescended or retractile testis, presence of varicocele, testicular volume, levels of serum follicle‐stimulating hormone, luteinising hormone, and testosterone, presence of Klinefelter syndrome and micro‐deletion of Y chromosome on sperm retrieval rates were evaluated. In 45.8% (n = 394) of the patients who underwent micro‐TESE, spermatozoon was adequately obtained. Multiple logistic regression analysis demonstrated that previous successful testicular biopsy (OR = 15.346; GA = 5.45–43.16; p < .001) and higher testicular volumes significantly increase sperm retrieval rate in micro‐TESE. The testicular volume cut‐off as 11 ml was found to be the most significant factor. Although currently testicular biopsy result is not being used as a diagnostic method, it is significantly associated with micro‐TESE result.     

Five years experiences with microinjection of testicular spermatozoa into oocytes in Hungary

The aim of the study was to summarize our five years experience (1996-2000) of testicular spermatozoa for intracytoplasmic sperm injection in Hungary. The influence of sperm count, maternal age, number of transferred embryos, and application of assisted hatching on outcome was investigated. Testicular spermatozoa were retrieved by microsurgical testicular sperm extraction. Samples were classified depending on the number of spermatozoa. Indication for testicular sperm extraction in conjunction with intracytoplasmic sperm injection was severe azoospermia or azoospermia combined with tubal origin infertility. Ovarian stimulation was carried out using an ultrashort protocol with GnRH agonist and gonadotrophin. Intracytoplasmic sperm injection was performed without PVP. Embryos were cultured for 48 or 72 h before embryo transfer. Indications for assisted hatching included elevated maternal age, increased zona thickness or at least two previous unsuccessful IVF cycles. Testicular spermatozoa were successfully retrieved in 218 out of 273 cases. Extreme low sperm count was found more frequently in cases of nonobstructive azoospermia. No significant differences were observed in fertilization rate (61.1% vs. 51.7%) or clinical pregnancy rate (29.0% vs. 26.7%) between patients with obstructive or nonobstructive azoospermia. Maternal age, number of transferred embryos and application of assisted hatching had a significant effect on outcome. A total of 55 clinical pregnancies were achieved, including 14 sets of twins, three sets of triplets and two sets of quadruplets. It is concluded that testicular sperm extraction is an efficient way of obtaining testicular spermatozoa, allowing not only successful fertilization by ICSI, but also freezing of testicular spermatozoa for use in subsequent cycles.     

Predictors for successful sperm retrieval of salvage microdissection testicular sperm extraction (TESE) following failed TESE in nonobstructive azoospermia patients

The purpose of this study was to observe the clinical utility and the possible determinants in predicting sperm retrieval of salvage microdissection testicular sperm extraction (mTESE) for nonobstructive azoospermia (NOA) patients with failed conventional TESE at their first attempts. A total of 52 NOA males underwent salvage mTESE were recruited in this study. Related data, including age, BMI, the presence of Klinefelter's syndrome and varicocele, cryptorchidism, mean testicular volume, hormonal profile (total testosterone (TT), follicle‐stimulating hormone (FSH), luteinising hormone (LH), inhibin B (INHB)), testicular histology and surgical duration, were collected and analysed. A multivariate logistic regression with likelihood ratio test revealed the following predictors of sperm retrieval: TT and testicular histology (chi‐square of likelihood ratio = 26.42, df = 4, p < .005). A formula was also established using multivariate regression analysis in predicting sperm retrieval probability. A predicted probability of more than 71% was determined of the formula as the cut‐off value in predicting sperm retrieval using receiver operating characteristics (ROC) analysis with a sensitivity and specificity 78.0% and 72.4% respectively. In conclusion, salvage mTESE is of clinical value in NOA males with failed TESE attempts, whereas the established formula could be useful in determining the proper salvage mTESE candidates.     

The predictive value of parameters of clinical presentations for sperm yield in patients with nonobstructive azoospermia receiving microdissection testicular sperm extraction

ObjectiveWe analyzed a cohort of nonobstructive azoospermia (NOA) patients receiving microdissection testicular sperm extraction (mTESE) to examine the relationship of sperm yield and the parameters of clinical presentations. We aim to identify the parameters that might positively predict a positive sperm yield after mTESE.Materials and methodsA total of 200 patients with NOA who had undergone mTESE were enrolled. Among them, 112 (56%) had received a prior testicular needle biopsy. Clinical data including physical findings, underlying genetic abnormalities, pathologic findings in needle biopsy, and sperm retrieval rate (SRR) during mTESE were reviewed and analyzed.ResultsThe pathological findings of prior needle biopsy demonstrate a predictive value of sperm yield during mTESE. Hypospermatogenesis had SRR of 93.3% during mTESE, early maturation arrest had SRR of 13.3%, late maturation arrest (LMA) had SRR of 66.7%, and Sertoli cell-only syndrome had SRR of 18.1%. Regarding parameters of clinical presentation, we found that SRR during mTESE was 85.7% for patients with hypogonadotropic hypogonadism, 60.0% for men with undescended testes (UDT) history, 50.0% for patients who had been exposed to chemotherapeutics due to malignancy of other organs, 100% for prior mumps infection, 50.0% for AZFc deletion, 50.0% for Klinefelter syndrome, and 33.3% for other sex chromosome-related abnormalities. No sperm was found in patients with AZFa or AZFb microdeletion. The consistency of histopathological findings between initial testis biopsy and mTESE was 77.7%. As much as 17.4% of cases had upgraded on spermatogenesis at later mTESE.ConclusionClinical presentations or phenotypes can be used as predictive factors for successful sperm retrieval during mTESE in patients with NOA. Hypogonadotropic hypogonadism and cases with UDT history have a higher chance of sperm retrieval. Initial testicular needle biopsy, if available, can provide valuable information about chances of sperm retrieval. Hypospermatogenesis predicts high sperm yield rate, and LMA can have best upgrade results of sperm yield after mTESE.     

Comparison of pregnancy and neonatal outcomes of intracytoplasmic sperm injection performed with frozen versus fresh testicular sperm

BackgroundIt remains controversial whether there is a difference in the prognosis of intracytoplasmic sperm injection (ICSI) using frozen or fresh testicular sperm in patients with obstructive azoospermia (OA). Moreover, in the available studies, few have tracked neonatal outcomes. This study aimed to compare the pregnancy and neonatal outcomes of ICSI using cryopreserved sperm versus fresh sperm collected by testicular sperm aspiration (TESA).MethodsA total of 317 OA patients treated with ICSI in a university affiliated hospital from January 2016 to December 2020 were included in this study. The participants were divided into two groups according to the type of sperm used for ICSI: frozen sperm group (n=154) and fresh sperm group (n=163). The pregnancy and neonatal outcomes of the two groups were compared.ResultsThe data produced by this study showed no significant statistical difference in the 2 pronuclei (2PN) fertilization rate, 2PN cleavage rate, high-quality blastocyst rate, and the average number of transferred embryos in the frozen and fresh sperm groups. Similarly, no difference was found in implantation rate, clinical pregnancy rate, multiple pregnancy rate, miscarriage rate, premature delivery rate, live birth rate, and gender ratio at birth (P>0.05). The average newborn birth weight was similar in both groups (2,932.61±728.40 vs. 3,100.32±515.64 g, respectively) (P>0.05). A higher incidence of low birthweight (LBW) newborns was found in the frozen sperm group (20.91% vs. 8.49%) (P<0.05). Multiple logistic regression analysis showed that LBW is related to single or twin pregnancies (P<0.01), but not sperm (frozen or fresh) (P>0.05). We further analyzed the twin and single pregnancies in the two groups separately, and found that the incidences of LBW were both similar (P>0.05). There was no difference in the Apgar scores at 1 min and 5 min after birth between the two groups (P>0.05).ConclusionsThe use of frozen testicular sperm by TESA was efficient for men with OA. There were similar pregnancy and neonatal outcomes following TESA-ICSI using frozen or fresh sperm in this retrospective study. Prospective investigations are needed for further validation.     

Freezing of testicular tissue as a minced suspension preserves sperm quality better than whole-biopsy freezing when glycerol is used as cryoprotectant

Frozen-thawed testicular spermatozoa have been used successfully for ICSI, especially in cases of obstructive azoospermia with normal spermatogenesis. Fewer attempts, however, have been made to check whether these rather immature spermatozoa, in a different environment with several other cell types present, have cryobiological requirements other than those of ejaculated spermatozoa. This is the reason why the freezing protocols and cryoprotectants (glycerol) used for freezing testicular tissue are based on experience with semen freezing. This study aimed to assess whether cryosurvival and/or motility was influenced by freezing of testicular tissue either as an intact biopsy or as a shredded tissue suspension, when glycerol was used as cryoprotectant. Freezing of testicular tissue as a suspension preserved motility (type B + C) significantly better than freezing of whole biopsies (9.2% vs. 4.0%). Similar observations have been made for vitality (39.3% vs. 25.4%). Centrifugation on 50% Percoll in order to remove the cryoprotectant resulted in a huge loss of spermatozoa (or late spermatids) and should therefore be especially avoided in cases of testicular failure. On the basis of these observations, mincing of the testicular biopsies before freezing may be advocated. Testicular spermatozoa seem to be better preserved when frozen in suspension, at least when slowly permeating glycerol is used as a cryoprotectant.     

The clinical and neonatal outcomes after stimulation of immotile spermatozoa using SperMagic medium

To evaluate the efficiency and safety of SperMagic medium on stimulating the immotile spermatozoa in testicular sperm extraction (TESE) and absolute asthenozoospermia, 96 patients with TESE and 106 patients with absolute asthenozoospermia were enrolled in this study. The motile spermatozoa were detected in 47 TESE patients and 68 absolute asthenozoospermia and these patients were assigned to control group. The immotile spermatozoa in 49 TESE patients and 34 absolute asthenozoospermia were stimulated with SperMagic medium. Patients were treated by standard intracytoplasmic sperm injection (ICSI). There were no significant differences in fertilisation, cleavage, implantation, pregnancy, live birth and neonatal outcomes. SperMagic medium does not increase incidence of adverse neonatal outcomes and is a reliable tool for selection of viable spermatozoa in ICSI.     

Outcome of repeated micro-surgical testicular sperm extraction in patients with non-obstructive azoospermia

Aim： To evaluate the outcome of repetitive micro-surgical testicular sperm extraction （mTESE） attempts in non-obstructive azoospermia （NOA） cases, in relation to patients＇ initial testicular histology results. Methods： A total of 68 patients with NOA in whom mTESE had been performed in previous intracytoplasmic sperm injection （ICSI） attempts were reviewed. Results： Among the 68 patients with NOA, the first mTESE yielded mature sperm for ICSI in 44 （64%） （Sp^＋）, and failed in the remaining 24 （36%） （Sp^-）. Following their first trial, 24 patients decided to undergo a second mTESE. Of these 24 patients, no spermatozoa were obtained in 5 patients, and Sp^＋ but no fertilization/pregnancy were achieved in 19. In these 24 cases, mTESE was successively repeated for two （n = 24）, three （n = 4） and four （n = 1） times. The second attempt yielded mature sperm in 3/5 patients from the Sp group and 16/19 patients from the Sp^＋ group. At the third and fourth trials, 4/4 and 1/1 of the original Sp^＋ patients were Sp^＋ again, respectively. Distribution of main testicular histology included Sertoli cell-only syndrome （16%）, maturation arrest （22%）, hypospermatogenesis （21%） and focal spermatogenesis （41%）. Overall, in repetitive mTESE, 24/29 （82%） of the attempts were finally Sp^＋. Conclusion： Repeated mTESE in patients with NOA is a feasible option, yielding considerably high sperm recovery rate. In patients with NOA, mTESE may safely be repeated one or more times to increase sperm retrieval rate, as well as to increase the chance of retrieving fresh spermatozoa to enable ICSI.     

Prediction of the extent of germ cell loss utilising a noninvasive spectroscopy method in rat testicular damage model

The aim of this study is to investigate the efficiency of elastic light single-scattering spectroscopy system, a noninvasive method, to acquire spectra during testicular biopsy from normal and damaged seminiferous tubules with various degrees of germ cell loss. Adult control rats and doxorubicin-injected rats to achieve seminiferous germ cell loss (for 10 days [10D], 20 days [D20], 30 days [D30], 40 days [D40], and 50 days [D50]) were used. Spectroscopic measurements were acquired utilising a single-fibre optical probe, and histopathology of the biopsied testicular tissue samples were compared. Time-dependent testicular damage comprising various degrees of seminiferous tubule degeneration after doxorubicin-administration was observed. In D30, D40 and D50 groups, where significant germ cell loss was identified, elastic light single-scattering spectroscopy system signals were well correlated with disturbed spermatogenesis where significant differences in spectral signals were obtained. Our findings indicate that the elastic light single-scattering spectroscopy system has the potential to enable instant imaging of spermatogenesis in rats and could also be useful in humans for clinical applications, such as to increase sperm recovery success during micro-TESE for men with nonobstructive azoospermia.     

Onco‐testicular sperm extraction: birth of a healthy baby after fertility preservation in synchronous bilateral testicular cancer and azoospermia

Testicular germ cell tumours (TGCT) represent 1%–1.5% of all male neoplasms, and they have the highest prevalence among men between 15 and 35 years old. Synchronous bilateral disease is a rare presentation, and the ratio of metachronous to synchronous bilateral disease is about 4 : 1. Several studies have suggested a correlation between male infertility and testicular cancer, with a 20‐fold increase in the incidence of testicular cancer in infertile patients compared with the general population. At the time of diagnosis, 50%–75% of patients with unilateral TGCT present with subfertility; almost 13% of the patients are azoospermic before treatment, and up to two‐thirds of patients become azoospermic following adjuvant cancer therapies. Therefore, fertility preservation should be considered in all oncological treatments. The only available option to preserve the reproductive potential in azoospermic patients with testicular cancer is to perform an onco‐testicular sperm extraction (onco‐TESE) before cancer treatment. In this paper, we describe a rare case of a patient with synchronous bilateral testicular cancer and azoospermia who was submitted to onco‐TESE, sperm cryopreservation, and which was followed by the delivery of a healthy baby after intracytoplasmic sperm injection (ICSI), emphasising the importance of fertility preservation in oncology patients.     

Follicle-stimulating hormone as a predictor for sperm retrieval rate in patients with nonobstructive azoospermia: a systematic review and meta-analysis

Noninvasive parameters for predicating sperm retrieval rate (SRR) are desirables. Follicle-stimulating hormone (FSH) has been an important predictor since the first years of testicular sperm extraction. Recent studies showed continuous interests in FSH, with both pros and cons. Thus, we conducted a meta-analysis to evaluate the diagnostic value of FSH as a predictor for patients with nonobstructive azoospermia (NOA) taking testicular sperm retrieval. Eligible diagnosis tests were identified from electronic databases (Cochrane Central Register of Controlled Trials, Medline, and EMBASE) without language restrictions. The database search, quality assessment, and data extraction were performed independently by two reviewers. The reference standard was the sperm retrieval result. Diagnostic value of FSH were explored by area under receiver operation characteristics (ROC) curve using Review Manager, version 5.1.0 (Cochrane Collaboration, Oxford, UK) and Meta-DiSc, version 1.4. Meta regression will be done if there is heterogeneity. Then, we find 11 tests including a total of 1350 patients met the inclusion criteria. Our pooled analysis showed that the area under ROC curve of FSH was 0.72 ± 0.04. Meta regression analyses showed that region and average age have an influence on the diagnostic value. FSH showed more diagnostic value with patients in East Asia and with younger patients. We concluded that FSH had moderate value in independently predicating SRR in men with NOA (area under curve >0.7). More detailed diagnosis tests should be anticipated in the future to confirm the diagnostic value of other noninvasive parameters.     

非梗阻性无精子症患者睾丸活检组织细胞悬液检查与病理组织学检查精子检出率的差异性研究

目的:比较非梗阻性无精子症(NOA)患者睾丸活检组织细胞悬液检查与病理组织学检查精子检出率的差异,探讨两种检查方法结果不一致时获取精子的可靠性及临床治疗方案的选择。方法:1 112例NOA患者接受睾丸精子抽吸术(testicular sperm extraction,TESE),睾丸活检组织分别进行细胞悬液检查和病理组织学检查。结果:两种检查方法结果一致率为92.63%,一致精子检出率为41.82%,一致精子未检出率为50.81%。Kappa分析表明两种检查方法的一致性强度属于最强。25例进入辅助生殖周期的细胞悬液检查发现精子而组织学检查未发现精子患者中,24例患者取卵日成功获取精子(取精成功率为96.0%)并实施卵胞质内单精子注射(ICSI),其治疗结局为8例临床妊娠(33.33%)、4例流产(16.67%)、12例未妊娠(50.0%)。结论:实施诊断性TESE时,采用睾丸活检组织的细胞悬液检查与组织病理学检查双重评估精子检出率的方法,结果一致率高并且迅捷、准确、可靠,为NOA患者进入辅助生殖周期时成功取到精子提供了保障。当两种检查方法结果不一致时,细胞悬液检查对临床治疗方案的选择指导意义更大。     

非梗阻性无精子症患者睾丸体积、生殖激素水平与睾丸穿刺取精结果的相关性研究

目的:探讨非梗阻性无精子症患者睾丸体积、生殖激素水平与睾丸穿刺取精术(TESA)结果的相关性,以及可用于预测TESA结果的睾丸体积、生殖激素水平的切点值,从而为非梗阻性无精子症患者进一步诊疗提供重要资料。方法:121例研究对象均为非梗阻性无精子症患者(NOA),测定其睾丸体积和生殖激素水平,并根据TESA结果分为无精子组和有精子组。结果:无精子组和有精子组的左侧睾丸体积(ml)、右侧睾丸体积(ml)、泌乳素(PRL,ng/ml)、卵泡刺激素(FSH,mIU/ml)、黄体生成素(LH,mIU/ml)、雌二醇(E2,pmol/L)、血清总睾酮(TT,nmol/L)水平分别为7.07±1.06和11.75±1.38、7.37±1.37和11.70±1.98、12.43±11.69和9.60±4.55、15.77±10.84和8.01±7.43、6.12±2.92和8.11±20.11、119.36±43.52和141.12±48.33、11.43±4.05和12.46±4.60。无精子组血清FSH和PRL水平平均值高于有精子组,并且有显著的统计学差异。虽然无精子组的睾丸体积平均数小于有精子组,但两组之间没有统计学差异。对于年龄、血清E2和TT水平,两组之间也没有统计学差异。利用ROC曲线优选的睾丸体积切点值为9 ml,此点其敏感性为93.8%/89.6%(左/右),特异性为100%/94.3%(左/右),睾丸体积ROC曲线的AUC为0.984/0.961(左/右),表明其诊断准确性较高;优选的血清FSH水平切点值为8.18 mIU/ml,此点其敏感性为71.2%,特异性为75.0%,FSH水平ROC曲线的AUC为0.743,表明其诊断准确性中等。结论:睾丸体积和FSH水平对于预测NOA患者TESA结果具有重要意义,并且睾丸体积诊断准确性明显优于FSH。     

不同穿刺针应用于睾丸穿刺取精术的比较

目的:比较三种不同穿刺针用于睾丸穿刺取精术(TESA)的效果和并发症,以助于临床选择。方法:先后使用1.6侧孔针、0.7蝶形针和1.2侧孔针对188例无精子症患者实施TESA,分别为102例、41例和45例。比较各自穿刺针数及1周内阴囊血肿、感染的发生率。结果:三种穿刺针术后1周内均未见明显并发症。采用1.6侧孔针进行1~3次穿刺,99例获得足够睾丸组织。0.7蝶形针获得满意组织,梗阻性无精子症(OA)患者需要(3.62±0.80)次穿刺,非梗阻性无精子症(NOA)患者需要(5.50±1.10)次。1.2侧孔针用于OA和NOA患者分别需要穿刺(1.95±0.69)次和(2.92±1.15)次。0.7蝶形针穿刺次数多于1.2侧孔针(P0.05)。NOA患者比OA更难穿刺获得睾丸组织(P0.05)。结论:OA和NOA患者以0.7蝶形针行TESA术,均需要比1.6和1.2侧孔针更多的穿刺针数,可能增加潜在并发症。拟行TESA的NOA患者,建议采用1.2侧孔针。     

不采用冷冻保护剂的玻璃化法与常规冷冻法对人精子冷冻复苏效果的比较

目的比较不采用冷冻保护剂的玻璃化法与常规冷冻法对人精子冷冻复苏的效果。方法将15份上游后的精液标本分别采用常规精子冷冻和不使用冷冻保护剂的冷冻环玻璃化法冷冻,比较精子复苏后的活力、形态及精子膜的完整性三项指标。结果冷冻前、前向活动精子百分率、正常精子形态百分率及精子膜完整率分别为(79±6.42)%、(34±9.36)%和(91±5.18)%;不采用冷冻保护剂的玻璃化法冷冻复苏后,三者分别为(42.20±8.35)%、(31.00±7.63)%和(50.00±9.34)%。常规冷冻法冷冻复苏后,三者分别为(38.00±15.80)%、(30.00±5.24)%和(47.00±13.67)%。冷冻前后前向活动精子百分率和精子膜完整率的差别有统计学意义,但两种冷冻方法相比差异无统计学意义。结论使用不加入冷冻保护剂的玻璃化方法冷冻人的精子是可行的,可取得与常规冷冻相同的效果。     

Kruger strict morphology and post-thaw progressive motility in cryopreserved human spermatozoa

The purpose of this prospective study was to evaluate Kruger strict morphology and conventional semen analysis in predicting cryosurvival and the progressive motility recovery rate of frozen spermatozoa. Our study included 56 semen samples with >10 million spermatozoa per ejaculate. The main outcome measures were conventional semen analysis, strict morphology analysis by the Kruger method, cryosurvival rate and post-thaw sperm motility. A significant reduction in sperm motility after cryopreservation was demonstrated. The freeze-thawing process caused a 66% reduction in rapid progressive motile spermatozoa, a 45% reduction in slow progressive motile spermatozoa and a 2% reduction in nonprogressive motile spermatozoa. The cryosurvival and progressive motility recovery rates were not correlated with parameters of conventional semen analysis, such as sperm concentration, motility, WHO morphology and total motile count, but the progressive motility recovery rate was significantly correlated with the percentage of spermatozoa exhibiting Kruger normal morphology (P = 0.028). The recovery rate of rapidly progressive motility was profoundly decreased compared with slow progressive motility following the frozen-thaw procedure of semen. Kruger strict morphology assessment was a better predictor of the progressive motility recovery rate following the freezing-thaw procedure than parameters of conventional semen analysis.     

Ideal culture time for improvement in sperm motility from testicular sperm aspirates of men with azoospermia

PURPOSE: The motility of testicular derived spermatozoa reflects viability and predicts success during intracytoplasmic sperm injection. Although improvements in sperm motility are seen after incubation for extended periods, no guidelines suggest duration or media use for optimal improvement in motility. MATERIALS AND METHODS: Between July 1999 and February 2005 testicular aspirations were performed on 95 men with azoospermia, including 51 with obstructive azoospermia and 44 with nonobstructive azoospermia. Sperm motility was determined at initial collection and following incubation for 24 or 48 hours in processing media or Ham's F10 + protein. A mixed regression model controlling for testis side, media and baseline motility was created to analyze the change in motility between 24 and 48 hours. RESULTS: Mean motility improved from 3% to 20% at 24 hours and 25% at 48 hours for OA cases and from 0% to 5% at 24 hours and 11% at 48 hours for nonobstructive azoospermia cases. The improvement in motility from 24 to 48 hours was significant for obstructive azoospermia cases (p = 0.001). While media was a nonsignificant factor in regression models, when patients were grouped into categories of motility change there was a significantly better response to F10 compared to processing media (p = 0.03). CONCLUSIONS: Incubation in processing media or Ham's F10 + albumin media improves sperm motility with significant improvement noted between 24 and 48 hours for obstructive azoospermia cases. Ham's F10 + albumin media may provide extra benefit for cases of nonobstructive azoospermia or nerve injury. These results suggest the ideal timing of oocyte retrieval for intracytoplasmic sperm injection correlates with 48-hour sperm incubation for obstructive azoospermia cases, and 24 hours for nonobstructive azoospermia and nerve injury cases.