Clinical outcome of cycles with oocyte degeneration after intracytoplasmic sperm injection

There are variant rates of oocyte degeneration after intracytoplasmic sperm injection (ICSI) among different patients. Oocyte degeneration after ICSI may reflect the cohort of oocyte quality and subsequent embryo development capacity and clinical outcome. This retrospective study analyzed 255 cycles with at least one degenerated oocyte after ICSI (degeneration group) and 243 cycles with no degenerated oocytes after ICSI (control group). Basic characteristics like female age, body mass index, duration of infertility, hormone (FSH, LH, E₂) levels on day 3 of menses, and primary infertility patient rate were similar between the two groups (p > 0.05). Total dose of gonadotropin and length of stimulation were also similar between the two groups (p > 0.05), but the degeneration group exhibited a more exuberant response to ovarian stimulation as reflected by more oocytes retrieved (p < 0.05). The number of 2PN embryos available and high quality embryos were similar between the two groups (p > 0.05), but the high quality embryo rate, early cleavage embryo rate, and available embryo rate were all statistically lower than the control group (p < 0.05). Embryo developmental kinetics seemed to be disturbed and embryo fragmentation rate increased in the degeneration group (p < 0.05). However, there was no statistical difference in the distribution of graded embryos transferred, and there were no statistical differences in the pregnancy rate, implantation rate, and abortion rate between the two groups (p > 0.05). We deduce that the presence of oocyte degeneration after ICSI may be associated with decreased embryo quality with embryo development kinetics disturbed. However, the clinical outcomes may not be affected if the premise that sufficient high quality degeneration group embryos are available for transfer.     

Comparison of glucose metabolism in in vivo- and in vitro-matured tammar wallaby oocytes and its relationship to developmental potential following intracytoplasmic sperm injection

Although marsupial oocytes undergo nuclear maturation in vitro, there is, at present, no indication of their developmental potential, largely owing to the lack of in vitro fertilisation and related technologies for marsupials. Glucose metabolism has proven a useful indicator of oocyte cytoplasmic maturation and developmental potential in several eutherian species. Therefore, the aims of the present study were to compare: (1) the rates of glycolysis and glucose oxidation in immature, in vitro-matured and in vivo-matured tammar wallaby oocytes; and (2) the metabolic rate of individual oocytes with their ability to form pronuclei after intracytoplasmic sperm injection. The rates of glycolysis measured in immature (2.18 pmol oocyte(-1) h(-1)), in vitro- matured (0.93 pmol oocyte(-1) h(-1)) and in vivo-matured tammar wallaby oocytes (0.54 pmol oocyte(-1) h(-1)) were within a similar range to values obtained in eutherian species. However, unlike the trend observed in eutherian oocytes, the glycolytic rate was significantly higher in immature oocytes compared with either in vivo- or in vitro-matured oocytes (P < 0.001) and significantly higher in in vitro-matured oocytes compared with in vivo-matured oocytes (P < 0.001). No relationship was identified between glucose metabolism and the developmental capacity of oocytes after intracytoplasmic sperm injection when assessed after 17-19 h. Oocytes that became fertilised (two pronuclei) or activated (one or more pronucleus) were not distinguished from others by their metabolic rates. Longer culture after intracytoplasmic sperm injection (e.g. blastocyst stage) may show oocyte glucose metabolism to be predictive of developmental potential; however, culture to the single-cell stage did not reveal any significant differences in normally developing embryos.     

辅助激活联合补救单精子胞浆注射在常规受精失败卵母细胞中的应用

目的:探讨辅助激活联合单精子胞浆注射(ICSI)技术在常规IVF受精失败周期中的应用。方法:15例常规IVF受精失败患者卵母细胞接受辅助激活联合ICSI处理(激活组),另15例常规IVF受精失败患者卵母细胞直接行补救ICSI处理(非激活组),记录和分析未受精卵母细胞经辅助激活联合ICSI处理后所获受精率、优质胚胎率、囊胚形成率及妊娠结局。结果:激活组(15周期,220枚未受精卵母细胞)正常受精率、卵裂率、优质胚胎率、囊胚形成率分别为50%(110/220)、90.9%(100/110)、33%(33/100)及15%(15/100),其中妊娠1例;非激活组(15周期,200枚未受精卵母细胞)正常受精率、卵裂率、优质胚胎率及囊胚形成率分别为85%(170/200)、92.9%(158/170)、9.5%(15/158)及0,无妊娠发生。在正常受精率上非激活组高于激活组,且存在显著性差异(P<0.05),而在优质胚胎率和囊胚形成率方面,激活组显著高于非激活组(P<0.05)。结论:辅助激活联合ICSI可以提高常规IVF受精失败周期的优质胚胎率及囊胚形成率进而改善妊娠结局。     

辅助生殖技术子代4岁龄运动发育结局分析

目的 探讨辅助生殖技术(assisted reproductive technology,ART)对子代远期运动发育结局的影响。方法 选取2009年1-12月在南京市妇幼保健院生殖医学中心进行卵胞浆内单精子注射(intracytoplasmic sperm injection,ICSI)的单胎活产子代18例为研究对象(ICSI),以年龄、性别、胎数为匹配条件,选取在南京市妇幼保健院出生的活产婴儿36例为对照组(NC),排除孕周<32周,体重<1 500 g,有宫内窘迫,出生窒息史者进行研究。利用儿童运动评估系列(Movement Assessment Battery for Children,M-ABC)量表和儿童感觉统合发展检核表(3~6岁)分别比较两组儿童4岁龄的运动及感觉统合能力发育结局。结果 体格发育:ICSI子代出生身长[ICSI:(48.72±2.52)cm,NC:(50.33±1.24)cm]、出生体重[ICSI:(3.04±0.68)kg,NC:(3.50±0.42)kg]均低于对照组,差异有统计学意义(P<0.05);4岁龄,两组的身高、体重及头围均无统计学差异;运动发育:两组在手灵巧度、目标/抓握运动及总分方面无统计学差异(P均>0.05);在平衡能力(ICSI:8.44±1.72,NC:10.92±3.18)方面,ICSI组能力低于对照组,差异有统计学意义(P<0.05)。感觉统合能力:两组在前庭平衡和大脑双分化情况、触觉防御、发育期运用障碍、视觉空间和形状感觉、重心、整体评估差异均无统计学意义(P均>0.05)。结论 通过细化评估内涵、扩大样本量和延长随访时间可以为辅助生殖子代远期发育结局的判断提供更有力的证据。     

不同来源精子行卵胞浆内单精子显微注射结局分析

目的 探讨不同来源精子行卵胞浆内单精子显微注射后的妊娠结局.方法 回顾分析218个卵胞浆内单精子显微注射周期,按精子来源分为射出精子组190个周期,附睾穿刺取精组12个周期,睾丸穿刺取精组16个周期,比较3组的受精率、卵裂率、优质胚胎率和临床妊娠率等指标.结果 睾丸穿刺取精组较射出精子组及附睾穿刺取精组的受精率、卵裂率高,经比较差异均有统计学意义(χ2值分别为15.27、10.83、7.87、4.25,均P<0.05);睾丸穿刺取精组与射出精子组及附睾穿刺取精组临床妊娠率比较差异均无统计学意义(χ2值分别为1.67、0.94,均P>0.05);3组间的流产率、畸形率均为0,差异均无统计学意义(均P>0.05).结论 附睾穿刺取精组及睾丸穿刺取精组来源精子行卵胞浆内单精子显微注射助孕可获得与射出精子组相似的临床妊娠率.     

卵胞浆内单精子显微注射技术的临床应用

目的:探讨男性因素不育症和常规体外受精(IVF)失败者采用卵胞浆内单精子显微注射(ICSI)治疗的临床效果.方法:对51例男性因素不育症和常规体外受精(IVF)失败者共计59个ICSI治疗周期进行了回顾性分析.结果:59个ICSI治疗周期共取卵596个,MⅡ期卵母细胞471个,MⅡ期卵母细胞正常受精率82.21%,卵裂率89.04%,临床妊娠19例,周期临床妊娠率为32.20%.结论:对男性因素不育症和常规体外受精(IVF)失败者ICSI是一种有效的治疗方法.     

卵胞浆内单精子注射后多原核合子发生的相关研究进展

多原核合子是常规体外受精-胚胎移植过程中的正常现象,卵胞浆内单精子注射(ICSI)避免了多精子受精造成的多原核合子,但仍有可能出现多原核合子.ICSI后多原核合子出现主要是由于第二极体排出异常造成,卵母细胞质量、男方精液情况以及ICSI操作技术本身都是多原核合子形成的影响因素.该文针对ICSI后多原核合子的产生机制、影响因素、妊娠结局及相应处理措施进行综述.     

精子DNA完整性与IVF/ICSI助孕结局的关系

目的:探讨精子DNA碎片与体外受精/卵胞浆内单精子注射(IVF/ICSI)结局的关系。方法:采用精子染色质扩散实验(SCD)对接受IVF/ICSI治疗的280例(IVF 208例、ICSI 72例)男性精液样本进行DNA碎片指数(DFI)检测。根据检测结果将精液样本划分为低DFI(25%)组和高DFI(≥25%)组,并比较两组的受精率、卵裂率、优胚率、可利用胚胎率及临床妊娠率。结果:无论选择何种受精方式,低DFI组和高DFI组的受精率、卵裂率、优胚率、胚胎利用率及临床妊娠率均无统计学差异(P0.05)。结论:DFI的高低对IVF/ICSI的各种指标及助孕结局的影响不大。     

Porcine embryos produced after intracytoplasmic sperm injection using xenogeneic pig sperm from neonatal testis tissue grafted in mice

Embryo development after homologous intracytoplasmic sperm injection (ICSI) with sperm from testis tissue xenografts from pigs or any other farm animal species has not been evaluated critically. Here, we report development of porcine embryos in vitro following ICSI with sperm retrieved from xenografted neonatal pig testis. Small pieces of testis tissue from newborn piglets were grafted under the back skin of castrated immunodeficient mice (n = 4) and the xenografts were collected 8 months after grafting. Spermatozoa were recovered by mincing of the grafted tissue. For comparison, testicular, epididymal and ejaculated spermatozoa were also collected from mature boars. Oocytes injected with xenogeneic spermatozoa were either fixed to determine fertilisation processes (n = 89 in five replicates) or allowed to develop in vitro (n = 143 in four replicates). Xenogeneic porcine spermatozoa were fertilisation competent (24% v. 58%, 68%, 62% or 0% for xenogeneic v. control testicular, epididymal and ejaculated spermatozoa or no spermatozoa, respectively) and embryos developed to the blastocyst stage (8% v. 22%, 27%, 25% or 0%, respectively). These results demonstrate that porcine spermatozoa derived from immature testis tissue xenografted into mice are fertilisation competent, albeit at a lower rate than testicular, epididymal or ejaculated spermatozoa from control boars, and support embryo development after ICSI.     

Effect of centrifugation and electrical activation on male pronucleus formation and embryonic development of porcine oocytes reconstructed with intracytoplasmic sperm injection

Oocyte centrifugation and electrical activation are commonly used in intracytoplasmic sperm injection (ICSI) of bovine and porcine oocytes, to facilitate visual identification of sperm release into the ooplasm and to support oocyte activation following injection with tail membrane-damaged sperm. The present study evaluated the necessity of these steps in porcine modified ICSI. In the first series of experiments, in vitro-matured gilt oocytes with or without centrifugation were injected with head membrane-damaged spermatozoa aspirated tail first. Oocytes without centrifugation exhibited a significantly higher normal fertilisation rate, defined as male pronucleus (MPN) and female pronucleus (FPN) formation and the presence of two polar bodies, than centrifuged oocytes (40% v. 9%, respectively; P < 0.05). The rate of MPN formation was significantly higher in uncentrifuged oocytes compared with centrifuged oocytes (48% v. 17%, respectively; P < 0.05). The rates of survival, cleavage, blastocyst formation and total cell number in blastocysts did not differ between the two groups of oocytes. Next, the effect of electrical activation after ICSI on uncentrifuged oocytes injected with head membrane-damaged spermatozoa was determined. No significant differences were observed in the rate of MPN formation in sperm-injected oocytes regardless of electrical activation. However, the survival rates of sperm-injected or control oocytes without electrical activation were significantly higher than those of sperm-injected or control oocytes with electrical activation (88% and 84% v. 77% and 64%, respectively; P < 0.05). The cleavage rates of sperm-injected oocytes were significantly higher than those of control oocytes, regardless of electrical activation (77% and 81% v. 47% and 61% in sperm-injected and control oocytes with or without electrical activation, respectively; P < 0.05). Although development to blastocysts was similar in all experimental groups, the total cell numbers in blastocysts from control oocytes were significantly higher than those in sperm-injected oocytes, regardless of electrical activation (40 and 44 v. 22 and 26 in control and sperm-injected oocytes with or without electrical activation, respectively; P < 0.05). In conclusion, the present study clearly demonstrated that oocyte centrifugation before sperm injection is not beneficial to normal fertilisation and that electrical activation is not necessary in the modified porcine ICSI.     

附睾精子抽吸术结合卵细胞浆内单精子注射治疗阻塞性无精子症所致男性不育的研究

目的:探讨附睾精子抽吸术(ESA)结合卵细胞内单精子注射(ICSI)技术治疗阻塞性无精子症所致男性不育的治疗效果。方法:选择2002年1月~2003年12月到我院治疗不育症确诊为阻塞性无精子症男性不育患者,采用ESA方法吸取男性附睾液,分离精子用于ICSI;同时按常规体外受精-胚胎移植方法(IVF-ET),采用GnRH-α+FSH/hMG+hCG促排卵方案对女性进行促排成熟卵细胞(M II)用于显微注射授精,受精卵体外培养3 d后移植回子宫内。结果:采用MESA结合ICSI技术治疗32周期阻塞性无精子症所致不育的夫妇,所获成熟卵(M II)162个,受精率66.28%,卵裂率62.21%,临床妊娠率31.20%。结论:采用ESA结合ICSI技术治疗阻塞性无精子症所致男性不育获得良好的效果,该方法为阻塞性无精子症男性不育患者提供了一种快速、方便、无痛、有效的治疗方法。     

Effects of bull, sperm type and sperm pretreatment on male pronuclear formation after intracytoplasmic sperm injection in cattle

This study investigated the effects of the bull, sperm type (dead, immotile or motile) and sperm pretreatment (i.e. mechanical (tail-cutting or tail-scoring) or chemical (heparin, heparin + caffeine, calcium ionophore A23187 or dithiothreitol)) on male pronuclear formation after intracytoplasmic sperm injection (ICSI) in cattle. Three experiments were conducted. In Experiment 1, spermatozoa from three bulls (A, B and C) were used for both ICSI and in vitro fertilization (IVF). The results were that sperm from bull B yielded a higher penetration/male pronuclear formation rate than that of bull C when used for IVF (89.6% v 25.6%, P<0.01). However, when injected into oocytes by ICSI, sperm from bull C had a higher male pronuclear formation rate than that of bull B (34.6% v. 16.1%, P<0.05). The effects of sperm type and mechanical pretreatment were examined in Experiment 2. No significant difference was found in the male pronuclear formation rate when the three types of sperm were injected into oocytes. Tail-scored sperm achieved a higher male pronuclear rate than that of non-mechanically treated ones (38.2% v. 13.2%, P<0.005). In Experiment 3, chemical pretreatments were tested and compared. Higher male pronuclear rates, compared with the control, were obtained when sperm were pretreated with heparin + caffeine, calcium ionophore A23187 and dithiothreitol (48.2%, 62.5% and 64.5% v. 25.0%, P<0.05, 0.005 and 0.005, respectively). These results indicate that (1) there is a bull variation in male pronuclear formation with ICSI, and the male pronuclear rate by ICSI is not coincident with the results by IVF, (2) immobilization of a spermatozoon by tail-scoring before ICSI can improve the formation of the male pronucleus, and (3) an appropriate chemical pretreatment of spermatozoa is necessary to achieve a higher rate of male pronuclear formation.     

利用附睾或睾丸精子行卵胞浆内单精子注射治疗阻塞性无精子症

目的 回顾性分析38例阻塞性无精子症（OA）患者利用附睾或睾丸精子行卵胞浆内单精子注射（ICSI）的治疗结局。方法OA患者通过经皮附睾精子抽吸术（PESA）或睾丸切开取精术（TESE）获得精子行ICSI，评估受精率及临床妊娠率，以精液精子行ICSI组为对照。结果OA组38例共41个治疗周期，受精率和临床妊娠率分别为73．3％和53．6％，精液精子组31例33个治疗周期受精率和临床妊娠率分别为75．1％和48．4％，两组间比较，受精率和临床妊娠率差异无统计学意义（P〉0．05）。OA组共妊娠22例，已分娩13例，流产6例，继续妊娠3例；精液精子组共妊娠16例，已分娩10例，流产1例，继续妊娠5例。结论采用附睾／睾丸精子行ICSI是治疗男性阻塞性无精子症的有效方法。     

Testicular sperm extraction (TESE) with intracytoplasmic sperm injection (ICSI) now allowed in the Netherlands

After a moratorium of more than 10 years, it is again possible in the Netherlands to perform testicular sperm extraction (TESE) in combination with intracytoplasmic sperm injection (ICSI). Under the strict conditions of a research protocol, couples, of whom the man has a non-obstructive azoospermia, have the chance to parent their own offspring. The described procedure is an important step in the careful introduction of this new reproductive technique in the Netherlands.     

卵胞浆内单精子注射子代4岁龄智能发育情况分析

目的 分析卵胞浆内单精子注射(ICSI)子代4岁龄智能发育情况。方法 选取2009年1月-2009年12月在南京市妇幼保健院进行卵胞浆内单精子注射的单胎活产子代作为研究对象(ICSI,n=18例),以年龄、性别为匹配条件,选取该院出生的自然受孕单胎活产婴儿为作对照组(SC,n=36例);排除标准:孕周<32周,体重<1 500g,有宫内窘迫,出生窒息史者。利用中国韦氏学前儿童智力量表(C-WPPSI)分别比较两组儿童4岁龄的智力发育水平。结果 比较两组儿童4岁龄智能发育水平后发现,韦氏智力测验中ICSI子代总智商、言语智商及操作智商均值在正常范围,但得分均低于SC组(总智商:ICSI 100.67±11.45,SC 115.33±10.82;言语智商:97.72±14.21,111.72±12.40;操作智商:104.83±7.87,116.50±10.68),两者差异具有统计学意义(P<0.05)。进一步分析两组智力异常的发生率,发现ICSI子代图画填充项目量表分<7的人数明显高于SC组(ICSI 4/18,SC 0/36,χ²=8.64,P<0.05)。结论 有限的样本量结果提示,ICSI子代4岁龄智能发育在正常范围,但韦氏智力测验IQ值低于SC组。ICSI子代远期智能情况需要通过更广泛、长期、系统的随访跟踪才能得出更为详实的结论。     

利用附睾或睾丸精子行卵胞浆内单精子注射治疗阻塞性无精子症

目的回顾性分析38例阻塞性无精子症(OA)患者利用附睾或睾丸精子行卵胞浆内单精子注射(ICSI)的治疗结局。方法OA患者通过经皮附睾精子抽吸术(PESA)或睾丸切开取精术(TESE)获得精子行ICSI,评估受精率及临床妊娠率,以精液精子行ICSI组为对照。结果OA组38例共41个治疗周期,受精率和临床妊娠率分别为73.3%和53.6%,精液精子组31例33个治疗周期受精率和临床妊娠率分别为75.1%和48.4%,两组间比较,受精率和临床妊娠率差异无统计学意义(P>0.05)。OA组共妊娠22例,已分娩13例,流产6例,继续妊娠3例;精液精子组共妊娠16例,已分娩10例,流产1例,继续妊娠5例。结论采用附睾/睾丸精子行ICSI是治疗男性阻塞性无精子症的有效方法。     

短时受精失败后早期补救卵胞浆内单精子注射的最佳窗口期

目的探讨短时受精失败后不同时间进行早期补救卵胞浆内单精子注射(ICSI)对胚胎发育潜能及妊娠结局的影响。方法回顾性分析2013年7月-2016年12月在该中心受精失败后按不同时间补救ICSI的患者资料,共97周期。按补救时间分为A组(短时受精后4~5 h,35周期)、B组(短时受精后5.1~6 h,36周)和C组(短时受精后6.1~8 h,26周期),比较各组间的受精、胚胎发育情况和妊娠结局。结果受精参数比较差异均无统计学意义(均P0.05);补救ICSI时间越早,优质胚胎率、胚胎利用率、胚胎种植率和临床妊娠率越高,A组的优质胚胎率和胚胎种植率显著高于B、C两组(P0.05),B组的优质胚胎率显著高于C组(P0.05),A组的胚胎利用率显著高于C组(P0.05);补救ICSI时间越早,周期取消率、生化妊娠率和流产率呈降低趋势,但差异无统计学意义(P0.05)。结论在卵母细胞老化前,第2极体排出后,选择最佳补救ICSI的窗口期有利于改善短时受精失败后胚胎质量,提高胚胎利用率,从而提高胚胎种植率。     

卵胞浆内单精子注射术后卵巢妊娠1例报道及文献复习

<正>1病例资料患者,女,31岁,因体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)术后30 d,下腹痛1 d于2018年6月30日入院,患者因丈夫极度少弱精子症于广东省计划生育专科医院行卵胞浆内单精子注射(intracytoplasmic sperm injection,ICSI)助孕,使用短     

体外受精失败后不同时间行补救性卵细胞浆单精子显微注射的对比分析

目的 通过对常规体外授精(IVF)失败(未观察到第二极体)后不同时间行补救性卵细胞浆单精子显微注射(ICSI)的临床资料比较,探讨补救性ICSI的最佳时间. 方法 回顾性分析常规体外授精(IVF)失败后6 h(早期组)与22 h(传统组)行补救性ICSI的临床资料,比较两组的受精率、卵裂率、优胚率、种植率和妊娠率. 结果 早期组受精率、种植率、临床妊娠率分别为74.6%(56/75)、37.5%(9/24)、53.8%(7/13);传统组受精率为63.0%(29/46)、种植率和临床妊娠率均为0.早期组7例妊娠中4例单胎,3例流产;早期组和传统组受精率比较,差异无统计学意义,妊娠率和种植率比较,差异有统计学意义(P<0.05); 结论 早期补救性ICSI在受精率、种植率和临床妊娠率上可能优于传统补救性ICSI.     

染色体多态性与单精子卵泡浆内注射辅助生殖妊娠结局的关系

目的:探讨染色体多态性与单精子卵泡浆内注射(ICSI-ET)辅助生殖的妊娠结局的关系。方法:对拟行IC-SI助孕患者进行常规外周血淋巴细胞培养,采用G显带技术,通过染色体的核型分析,比较染色体多态性组与染色体核型正常组患者的临床妊娠率、早期流产率和种植率。结果:染色体多态性组与正常组相比,胚胎种植率(29.4%vs 26.8%)、临床妊娠率(46.3%vs 50.8%)和早期流产率(10.5%vs 8.2%)均无统计学差异(P>0.05)。结论:染色体多态性对于ICSI-ET助孕患者的种植率、临床妊娠率、早期流产率无明显影响。