经皮睾丸精子抽吸术治疗无精子症的研究

目的探讨经皮睾丸精子抽吸术(PTSA)获取睾丸精子结合卵胞浆内单精子注射术(ICSI)治疗梗阻性和非梗阻性无精子症,使之获得亲生子女.方法对121例因男性梗阻性及非梗阻性无精子症患者进行诊断性穿刺,均证实有精子后进行119个周期PTSA ICSI治疗.结果共获卵子1514个,成熟卵985个,胚胎741个,平均每例6.23个胚胎,总受精率74.4%,卵裂率97.6%;共移植114个周期和冷冻胚胎移植5个周期,平均移植2.86个胚胎,B超证实临床妊娠48例,临床妊娠率40.3%.结论采用PTSA技术获取的睾丸精子进行ICSI是治疗梗阻性及非梗阻性无精子症的一种安全、简单、有效的方法.     

冷冻复苏的睾丸精子对ICSI助孕结局的影响

目的探讨冷冻复苏的睾丸精子对ICSI助孕结局的影响。方法回顾性分析2014年3月至2017年5月因男方梗阻性无精子症(OA)或非梗阻性无精子症(NOA)在我院采用细针睾丸穿刺抽吸精子行ICSI助孕的209个周期的临床资料,新鲜睾丸精子(新鲜睾丸精子组)用于174个周期,冷冻复苏的睾丸精子(冻精组)用于35个周期。比较两组的一般资料、受精率、卵裂率、优质胚胎率、临床妊娠率以及种植率。结果两组的一般资料比较无统计学差异(P0.05)。冻精组与新鲜睾丸精子组行ICSI助孕后的受精率、卵裂率、优质胚胎率、临床妊娠率及种植率比较均无统计学差异(P0.05)。结论冷冻复苏的睾丸精子对ICSI助孕结局并无显著影响。     

冷冻睾丸精子行卵胞浆内单精子注射临床结局探讨

目的探讨冷冻睾丸精子行卵胞浆内单精子注射(ICSI)对无精子症患者临床结局的影响。方法回顾性分析2017年1月至2018年12月在我院生殖中心经皮睾丸穿刺取精(TESA)行ICSI助孕的213个授精周期的临床资料。按精子来源分为两组,TESA冻精组(观察组):采用TESA冷冻精子,共114个周期;TESA组(对照组):采用新鲜TESA精子,共99个周期。比较两组的胚胎发育情况和临床结局。结果 TESA冻精组正常受精率(2PN率)显著低于TESA组,差异具有统计学意义(64.26% vs 73.89%,P0.01)。两组的2PN卵裂率、可用胚胎率、优质胚胎率和囊胚形成率差别无显著性(P0.05)。两组的临床妊娠率、胚胎种植率、多胎妊娠率、异位妊娠率、早期流产率和分娩率差异没有统计学意义(P0.05)。结论冷冻睾丸精子行ICSI的临床结局与新鲜睾丸精子总体相似。但需要注意冷冻睾丸精子可能降低ICSI后正常受精率。     

新鲜附睾精子与冷冻复苏后的附睾精子临床结局比较

目的探讨经皮附睾穿刺取精术（PESA）获得精子经冷冻复苏后行卵胞浆内单精子注射术（ICSI）的临床效果。方法将采用新鲜附睾精子的94例患者作为新鲜组对照,冷冻复苏后附睾精子的92例患者作为冻精组,比较二组的受精率,可利用胚胎率及妊娠率。结果新鲜组与冻精组相比受精率,可利用胚胎率及妊娠率无显著性差异（75.04%vs78.4%,78.3%vs81.3%,47.87%vs44.44%P〉0.05）。结论PESA精子经冷冻后有很好的复苏率（97.94%）,结合ICSI可得到与使用新鲜PESA精子同样的临床结局,可作为梗阻性无精症的治疗方法。     

显微取精术在非梗阻性无精子症患者中的临床应用及结局

目的观察睾丸显微取精术在非梗阻性无精子症患者中的临床应用及其结局。方法17例患者在配偶同步促排卵取卵前一天行睾丸显微切开取精术,获得精子患者行卵胞浆内单精子显微注射(ICSI)助孕,分析其受精率、可用胚胎率和临床妊娠结局。结果17例患者显微取精,13例成功获取精子,精子获得率为76.5%。这13对夫妇ICSI受精率为74.8%,可用胚胎率49.5%。3例新鲜周期移植,全部成功妊娠,1例已分娩1健康婴儿,另外2例持续妊娠中。其余10例行全胚冷冻,4例分别于胚胎冻存3个月后行解冻复苏移植,3例成功妊娠,2例已分娩(其中一例为非嵌合型Klinefelter综合征患者),另外1例持续妊娠中。结论睾丸显微切开取精术是使NOA患者和非嵌合型Klinefelter综合征患者成功获得自己遗传学子代的有效方法,有较高的精子获得率,联合ICSI技术、全胚冷冻-复苏移植可以获得较高的临床妊娠率。     

不同来源精子对ICSI助孕结局的影响

研究不同来源的精子对于ICSI辅助生殖助孕结局的影响。对本中心115例卵胞浆内单精子显微注射（ICSI）治疗周期进行回顾性分析,比较分别使用射出精液来源精子以及睾丸、附睾穿刺来源精子行ICSI后,受精率、卵裂率、优质胚胎率以及妊娠率、种植率等各项指标。精子来源的不同造成了受精率的显著差异,一旦受精完成,随后胚胎的发育及种植将更多的依赖于患者的年龄。     

补救ICSI的妊娠结局分析(附补救后冷冻胚胎复苏妊娠一例)

目的对体外受精(IVF)完全失败的周期及时补行卵细胞浆内单精子显微注射(rescue ICSI)后的临床妊娠结局的探讨和分析。方法IVF受精失败后及时将MⅡ卵母细胞进行补救ICSI。结果补行ICSI后10周期中移植9个周期19个胚胎,妊娠1例双胎,种植为10.5%;有2个周期有冷冻胚胎并行复苏胚胎移植,单胎妊娠1例。补救ICSI累计移植胚胎25个,种植3个累计种植率为12.0%,但2例均以流产告终。结论补救ICSI妊娠率低、流产率高与卵子过度老化有关,冷冻复苏补救ICSI的胚胎,可以消除胚胎发育与子宫内膜发育不同步的因素。在IVF受精率低的周期中可以将未受精的卵母细胞补行ICSI后行胚胎冷冻-复苏移植,可以增加取卵周期的妊娠率。     

关于提高单精子显微注射技术妊娠率的方法探讨

目的　探讨提高单精子显微注射技术 (intracytoplasmicsperminjection ,ICSI)妊娠率的方法。方法　采用胚胎操作箱进行卵的操作 ,应用人工血清代用品代替自体血清 ,多精子制动分次注射 ,分析其得卵率 ,卵裂率及妊娠率。结果　得卵率 15 5个 /周期 ,受精率 82 2 % ,优质胚胎占 80 6 % ,临床妊娠 6例 ,妊娠率 40 %。结论　采用胚胎操作箱进行卵的操作 ,应用人工血清代用品代替自体血清 ,多精子制动分次注射 ,可以提高妊娠率     

精子DNA碎片对IVF/ICSI结局的影响

目的探讨精子DNA碎片对体外受精/卵胞浆内单精子注射(IVF/ICSI)结局的影响。方法回顾性的分析了行IVF/ICSI治疗的725例不孕夫妇(IVF 502例、ICSI 223例)的临床资料。根据精子DNA碎片指数(DFI)分为正常组(DFI10%)、轻度DFI组(10%≤DFI20%)和重度DFI组(DFI≥20%),分别比较了IVF和ICSI中三组的受精率、卵裂率、可用胚胎率、优质胚胎率和临床妊娠率。结果在IVF周期中,DFI对受精率、卵裂率、可用胚胎率、优质胚胎率和临床妊娠率均无影响(P0.05);在ICSI周期中,DFI对受精率和卵裂率无影响(P0.05),但可用胚胎率和优质胚胎率三组之间有显著性差异(P0.05),临床妊娠率虽然随着DFI的增高而下降,却无显著相关性(P0.05)。结论精子DNA碎片对IVF结局基本无影响,但与ICSI结局中的可用胚胎率和优质胚胎率呈负相关,这可能与受精过程中IVF周期中卵子对精子有自然选择的过程,而ICSI周期中则是人为的选择精子有关。     

精子DNA碎片指数对冷冻胚胎移植临床结局的影响

目的通过评估精子DNA碎片指数(DFI),探讨精子DFI对冷冻胚胎移植结局的影响。方法采用改良精子染色质扩散实验(SCD)对530例接受体外受精(IVF)和153卵细胞浆单精子注射(ICSI)的男方进行精子DFI检测,并根据精子DFI值将上述IVF患者分组(DFI≤30%,DFI30%),根据单因素方差分析统计方法分析各组IVF受精率、卵裂率、优质胚胎率和冷冻胚胎移植的妊娠结局的差异。结果精子碎片指数DFI≤30%组在临床妊娠率,生化妊娠率和流产率与DFI30%组相比没有显著变化。在精子DNA碎片指数≤30%的ICSI组,囊胚形成率显著高于DFI30%组。结论精子染色质扩散试验(SCD)方法测定的精子DNA碎片指数与冷冻胚胎移植(FET)结局无关,但是与ICSI周期的囊胚形成率有关。     

The results of 154 ICSI cycles using surgically retrieved sperm from azoospermic men

BACKGROUND: The effects of source of sperm, aetiology and sperm cryopreservation on ICSI cycles in azoospermic men were evaluated. The effect of aetiology of azoospermia on embryo development was also assessed. METHODS: This study was a retrospective analysis of 154 cycles (91 couples) using surgically retrieved sperm. Outcome measures were fertilization rate (FR), implantation rate (IR), and clinical pregnancy rate (CPR) and livebirth rate (LBR) per transfer. RESULTS: Our data demonstrated similar outcome between the use of epididymal or testicular sperm in men with obstructive azoospermic (OA). FR and IR were significantly lower (P < 0.05) using sperm from men with non-obstructive azoospermic (NOA), but although pregnancy outcome appeared lower, this did not reach statistical significance (P = 0.08). Cryopreservation of epididiymal sperm did not alter outcome, but the use of frozen-thawed testicular sperm did demonstrate a lower FR, with no statistical difference in IR or pregnancy outcome. Embryos derived from NOA sperm had impaired development beyond day 2 post-oocyte retrieval (OA, 44% <5 cell; NOA, 71% <5 cell; P = 0.002). CONCLUSIONS: The use of sperm from men with NOA significantly affects fertilization and implantation in ICSI cycles. The use of frozen-thawed testicular sperm affects fertilization rate without significantly altering pregnancy outcome. The use of such data on which to base clinical decisions needs to be supported by the meta-analyses of previous reports.     

Fertilization, pregnancy and embryo implantation rates after ICSI with fresh or frozen-thawed testicular spermatozoa

This study aimed to determine whether fertilization and implantation rates after intracytoplasmic sperm injection (ICSI) with fresh or frozen-thawed testicular spermatozoa were comparable. Between 1 January 1996 and 31 December 1996, 65 ICSI cycles with testicular spermatozoa and 35 cycles with frozen-thawed testicular spermatozoa were carried out. In 50 out of 65 ICSI cycles, testicular spermatozoa could be retrieved and in 34 out of 35 cycles carried out with frozen-thawed testicular spermatozoa, motile spermatozoa could be recovered. The fertilization rate after ICSI with frozen-thawed testicular spermatozoa was significantly lower (71.1%; P < or = 0.008) than with fresh testicular spermatozoa (79.3%). The pregnancy rate was similar for both groups (38.2 and 26.5 %). The implantation rate per transferred embryo, however, was significantly lower in the frozen-thawed rather than in the fresh testicular sperm group (9.1 versus 24.6%; P = 0.001). The live birth rate per transferred embryo was also higher in the group in which fresh testicular spermatozoa were used (18.8 versus 7.9% P = 0.043). This retrospective study shows that is possible to achieve a high fertilization rate after ICSI with both fresh and frozen-thawed testicular spermatozoa but implantation and live birth rates per transferred embryo, however, are significantly lower after ICSI with frozen-thawed than with fresh testicular spermatozoa.      

Pregnancy with frozen-thawed and fresh testicular biopsy after motile and immotile sperm microinjection, using the mechanical touch technique to assess viability

BACKGROUND: There are few reports of pregnancy using immotile sperm, and none using a purely mechanical assessment of viability. METHODS: In this pilot study, we retrospectively analysed 66 cycles in 61 patients with determinant male factor, recording rates of fertilization, implantation, normal pregnancy and take-home babies achieved with ICSI. Sperm selection was based on morphologically normal appearance under the inverted microscope. Viability of immotile spermatozoa was assessed by the mechanical touch technique to observe tail flexibility and tail shape recovery. RESULTS: Of 17 ICSI cycles using frozen-thawed testicular sperm, six microinjected with immotile and 11 with motile sperm, we achieved fertilization rates of 65.7 and 74.3%, respectively, and five pregnancies (two and three, respectively). Of 49 ICSI cycles using fresh testicular sperm, 10 microinjected with immotile and 39 with motile sperm, we achieved fertilization rates of 73.4 and 64.4%, respectively, and 12 pregnancies (three and nine, respectively). CONCLUSIONS: Immotile (fresh and frozen-thawed) testicular sperm of normal morphological appearance can be used to achieve clinical pregnancy with ICSI. Our results strongly suggest that immotile sperm viability can be assessed by the mechanical touch technique.     

The outcome of intracytoplasmic injection of fresh and cryopreserved epididymal spermatozoa from patients with obstructive azoospermia--a comparative study

The aim of our study was to compare the outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) or microepididymal sperm aspiration (MESA) from patients with obstructive azoospermia. A retrospective analysis of consecutive ICSI cycles was performed, comparing the outcome in 24 patients with obstructive azoospermia undergoing surgical sperm aspiration by MESA (7 cycles) or PESA (17 cycles). In 23 of 24 patients, excess spermatozoa were cryopreserved. Following thawing, 21 ICSI cycles were performed (11 cycles after MESA, 10 after PESA). No statistically significant differences were noted in all parameters examined in ICSI cycles with fresh or cryopreserved spermatozoa from the same patients. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the rates of two-pronuclear fertilization (56% versus 53%), embryo cleavage (90% versus 86%), implantation (10% versus 14%), clinical pregnancy per embryo transfer (32% versus 37%) and delivery/ongoing pregnancy rate (27% versus 26%) were not statistically different. The cumulative ongoing pregnancy rate per sperm retrieval procedure was 46%, respectively. We conclude that the clinical outcome of ICSI with fresh and frozen-thawed spermatozoa after retrieval by PESA was similar to that by MESA. Epididymal sperm cryopreservation in patients with obstructive azoospermia is feasible and efficient using a simple freezing protocol and should be offered to optimize the yield of pregnancies achieved following such procedures.      

Outcome of in-vitro culture of fresh and frozen-thawed human testicular spermatozoa

The effect of in-vitro culture on the motility and morphology of fresh and frozen-thawed human testicular spermatozoa obtained from obstructive azoospermic patients and on the motility of testicular spermatozoa obtained from non-obstructive azoospermic patients was evaluated. The outcome of intracytoplasmic sperm injection (ICSI) with fresh and frozen-thawed human testicular spermatozoa was studied. The results showed that significant improvement of sperm morphology and motility was observed in culture of fresh (n = 17) and frozen-thawed (n = 15) testicular sperm samples obtained from patients with obstructive azoospermia. The motility of cultured testicular spermatozoa reached a peak at 72 h without the need for special media. In six of 20 samples obtained from patients with non-obstructive azoospermia, improvement of sperm motility was observed. When only non-motile testicular spermatozoa were cultured, they all remained non-motile (n = 9). In patients with obstructive azoospermia, fertilization rates of 80 and 81% were obtained using ICSI with fresh and frozen-thawed testicular spermatozoa respectively. Clinical pregnancies were observed in four out of nine patients with fresh testicular spermatozoa and two out of five patients after using frozen-thawed spermatozoa. When fresh testicular spermatozoa obtained from patients with non-obstructive azoospermia were used for ICSI, the fertilization rate was 68% and two out of seven patients achieved clinical pregnancies. In conclusion, the morphology and motility of fresh and frozen-thawed testicular spermatozoa in patients with obstructive azoospermia can be significantly improved after in-vitro culture. The outcome of in-vitro culture of testicular spermatozoa in patients with non-obstructive azoospermia is unpredictable. In-vitro culture of non-motile testicular spermatozoa is not successful so far. The outcome of ICSI with fresh and with frozen-thawed testicular spermatozoa was similar.      

Predictive value of testicular histology in secretory azoospermic subgroups and clinical outcome after microinjection of fresh and frozen-thawed sperm and spermatids

BACKGROUND: A retrospective study was carried out on 159 treatment cycles in 148 secretory azoospermic patients to determine whether histopathological secretory azoospermic subgroups were predictive for gamete retrieval, and to evaluate outcome of microinjection using fresh or frozen-thawed testicular sperm and spermatids. METHODS: Sperm and spermatids were recovered by open testicular biopsy and microinjected into oocytes. Fertilization and pregnancy rates were assessed. RESULTS: In hypoplasia, 97.7% of the 44 patients had late spermatids/sperm recovered. In maturation-arrest (MA; 47 patients), 31.9% had complete MA, and 68.1% incomplete MA due to a focus of early (36.2%) or late (31.9%) spermiogenesis. Gamete retrieval was achieved in 53.3, 41.2 and 93.3% of the cases respectively. In Sertoli cell-only syndrome (SCOS; 57 patients), 61.4% were complete SCOS, whereas incomplete SCOS cases showed one focus of MA (5.3%), or of early (29.8%) and late (3.5%) spermiogenesis. Only 29.8% of the patients had a successful gamete retrieval, 2.9% in complete and 77.3% in incomplete SCOS cases. In total, there were 87 ICSI, 39 elongated spermatid injection (ELSI) and 33 round spermatid injection (ROSI) treatment cycles, with mean values of fertilization rate of 71.4, 53.6 and 17%, and clinical pregnancy rates of 31.7, 26.3 and 0% respectively. CONCLUSIONS: Histopathological subgroups were positively correlated with successful gamete retrieval. No major outcome differences were observed between testicular sperm and elongated spermatids, either fresh or frozen-thawed. However, injection of intact round-spermatids showed very low rates of fertilization and no pregnancies.     

Retrospective multicentre study on mechanical and enzymatic preparation of fresh and cryopreserved testicular biopsies

BACKGROUND: Isolation of sperm suitable for ICSI from fresh or frozen-thawed testicular sperm extraction (TESE) can be facilitated by mechanical or enzymatic processing of the samples. METHODS: A retrospective multicentre study was initiated to compare these two approaches. Eleven German centres provided data on their TESE cycles performed during the period 1996/1997. Quality of retrieved sperm, fertilization rates of injected oocytes, embryo quality, resulting pregnancy rates and evolution of pregnancies were evaluated. RESULTS: The percentage of cycles with at least some motile sperm available for injection was higher after mechanical preparation. Independent of the preparation method, fertilization rates were higher for motile compared with immotile sperm or elongated spermatids in all groups and in general higher for cryopreserved versus fresh samples. Embryo quality was significantly better after injection of motile sperm for all treatments and in particular after enzymatic versus mechanical processing of biopsies. Pregnancy rates were identical for embryos derived from sperm prepared mechanically or enzymatically from fresh or cryopreserved testicular samples. The abortion rate (32/172, 18.6%) and the rate of multiple implantations (32/140, 22.9%) were not different from results reported in the literature for ICSI using ejaculated sperm. CONCLUSION: In this retrospective multicentre study, no unequivocal advantage of one over the other preparation method could be identified in 839 ICSI cycles using testicular sperm from 549 patients.     

Different fertilization rates between immotile testicular spermatozoa and immotile ejaculated spermatozoa for ICSI in men with Kartagener's syndrome: case reports

We report two cases of infertility treatment in couples where males suffered from Kartagener's syndrome (KS) and a total absence of motile sperm in the ejaculate. A total of three ICSI cycles was carried out. In all cycles, viable ejaculated or testicular spermatozoa were selected using the hypo-osmotic swelling (HOS) test. Case 1: In the first ICSI cycle total fertilization failure occurred after using ejaculated spermatozoa. In the following cycle testicular spermatozoa were used for ICSI, resulting in 75% fertilized oocytes and a pregnancy. Case 2: In the same ICSI cycle 50% of the oocytes were injected with ejaculated and 50% with testicular spermatozoa. The fertilization rates were 44 and 56% respectively and high quality embryos were achieved in both groups. One single embryo derived from testicular sperm was transferred with a resulting singleton pregnancy. In conclusion, testicular sperm for ICSI seem to have reliable fertilization capacity in men with KS, while ejaculated sperm, even if tested viable, seem more unpredictable. HOS test for selection of viable sperm for ICSI is recommended when ejaculated as well as testicular sperm are used for ICSI.     

Should diagnostic testicular sperm retrieval followed by cryopreservation for later ICSI be the procedure of choice for all patients with non-obstructive azoospermia?

BACKGROUND: This was a retrospective study to determine if diagnostic testicular biopsy followed by cryopreservation should be the procedure of choice for all patients with testicular failure. METHODS: The first part of the study analysed 97 ICSI cycles scheduled with frozen-thawed testicular sperm for 69 non-obstructive azoospermia (NOA) patients. The second part focused on a subgroup of 32 patients who underwent 42 ICSI cycles with frozen and 44 cycles with fresh testicular sperm. Sperm characteristics, fertilization, embryo quality, pregnancy and implantation rates were evaluated. RESULTS: Part I: The average time needed to find sperm was 113 min per cycle and 17 min per individual sperm. Fertilization rate, embryo transfer rate, ongoing pregnancy and implantation rates were 58.4%, 83%, 20.8% and 11.3%, respectively. Part II: The search time per sperm was higher (P=0.016) in frozen (18 min) than in fresh suspensions (13 min). A higher embryo transfer rate was observed in fresh cycles than in frozen cycles (93.2% vs 76.2%, P=0.028). Fertilization, ongoing pregnancy and implantation rates were comparable for the two groups. CONCLUSIONS: Even in a programme with low-restrictive criteria for patient allocation and for sperm cryopreservation, diagnostic testicular biopsy followed by cryopreservation can be the procedure of choice for patients with testicular failure.