未成熟卵母细胞体外培养在体外受精中对卵巢低反应性者的补救性应用

目的:探讨未成熟卵母细胞体外培养(IVM)在体外受精(IVF)中对卵巢低反应性者的补救性治疗效果。方法:30例行IVF时表现为卵巢低反应者(实验组)在B超引导下经阴道行卵泡穿刺术,取卵后对不同期未成熟卵母细胞行IVM,再行胞浆内单精子注射(ICSI)受精;对成熟卵母细胞则进行常规受精,胚胎形成后行移植术。取同期进行ICSI周期治疗的63例作为对照组。结果:应用IVM行IVF-ET补救治疗的30个周期中有29个周期获胚胎移植,临床妊娠9例(31%),胚胎种植率18.47%,但该组未成熟卵培养成熟后的受精率、卵裂率均低于对照组;总妊娠率及胚胎种植率比常规ICSI组低。结论:IVM技术联合IVF为卵巢低反应性者提供了一种有效的补救措施,明显减少了IVF周期的取消率,提高了妊娠机会。     

人未成熟卵母细胞体外培养成熟、受精及胚胎移植

目的 应用卵母细胞体外成熟(IVM)技术帮助卵泡成熟障碍的不孕症患者获得妊娠及分娩。方法 接受未成熟卵IVM技术治疗者30例35个周期,其中多囊卵巢综合征14例,有卵巢过度刺激综合征病史6例,体外受精和胚胎移植(IVF-ET)周期中卵巢反应不良患者10例。设计卵巢刺激方案,采用含人成熟卵泡液的IVM培养液,建立未成熟卵母细胞的体外培养方法。结果 35个周期共取得未成熟卵母细胞203个,平均每个周期5．8个。培养后有156个卵母细胞排出第一极体,IVM率76．8％(156／203);在卵胞浆单精子显微注射(ICSI)12～18h后观察原核,正常受精率为76．9％(120／156);共有移植周期33个,获8例临床妊娠,妊娠率24％(8／33);有5例共7个婴儿出生。结论 IVM对一些卵泡发育和成熟障碍,特别是顽固性多囊卵巢综合征患者,是一种有效的辅助生育措施。人成熟卵泡液含有理想的自然促卵母细胞成熟的成分。     

卵母细胞体外成熟预防卵巢过度刺激综合征的应用

目的:探讨卵母细胞体外成熟预防卵巢过度刺激综合征(ovarian hyperstim- ulation syndrome,OHSS)的效果。方法:51例IVF长方案超排卵过程中出现OHSS早期征象时,按患者意愿分成两组,实验组于当天注射人绒毛膜促性腺激素(hCG)36h后取卵,经体外培养成熟行体外受精、胚胎移植;对照组则继续按常规用药方案及受精方案行胚胎移植。测定两组出现OHSS早期征象当日的E_2、统计FSH用药天数、用药量、受精率及临床妊娠率。结果:实验组与对照组在出现OHSS早期征象当日的血E_2水平、受精率、优质胚胎率、临床妊娠率无统计学差异;但实验组的FSH总用量、FSH用药天数、获卵数方面显著低于对照组(P<0.05)。实验组1例发生中度OHSS,对照组则发生3例重度OHSS和2例中度OHSS,重度OHSS发生率差异极显著(P<0.01)。结论:超排卵过程中,出现OHSS早期征象时改行卵母细胞体外成熟培养技术可有效地降低OHSS的发生率。     

未促排卵的未成熟卵母细胞体外培养成熟治疗不孕症40例分析

目的比较未促排卵的未成熟卵母细胞体外培养成熟(IVM)治疗多囊卵巢综合征(PCOS)和正常月经周期伴不孕患者的临床疗效。方法回顾性分析2004年8月至2005年1月温州医学院第一医院收治的40例不孕患者,接受54个周期的IVM治疗,其中难治性PCOS不孕患者26例35个周期(A组),月经周期正常且经其它辅助生育技术失败的不孕患者14例19个周期(B组),均未采用任何药物刺激。结果共有7个周期取消,其中A组1个,取消率为2.8%;B组6个,取消率为31.6%,差异有极显著性意义(P<0.01)。有胚胎移植的47个周期中,两组的平均获卵数、体外成熟率、正常受精率和卵裂率、平均移植胚胎数差异无显著性意义(P>0.05)。A组1例生化妊娠,15例临床妊娠,B组4例临床妊娠。两组每穿刺周期的临床妊娠率(42.9%、21.1%)差异有极显著性意义(P<0.01),每移植周期的临床妊娠率、种植率和多胎率差异无显著性意义(P>0.05)。结论未促排卵的IVM技术治疗不孕症,尤其对于难治性PCOS不孕更是一种有效的方法。     

未成熟卵母细胞体外成熟技术在多囊卵巢综合征不孕症治疗中的应用

目的 探讨未成熟卵母细胞体外成熟技术(IVM)在多囊卵巢综合征(PCOS)不孕症治疗中的价值.方法 2003年1月至2009年12月,安徽医科大学第一附属医院生殖医学中心采用IVM技术治疗PCOS不孕症患者157例,共162个周期(IVM组);选取同期常规体外受精(IVF)或单精子卵细胞胞质内注射(ICSI)治疗PCOS不孕症患者109例,共114个周期(对照组1),以及非PCOS不孕症患者106例,共106个周期(对照组2)作为对照.比较3组患者治疗、妊娠结局及新生儿情况.结果 IVM组、对照组1、对照组2患者尿hCG阳性率分别为35.7％(56/157)、42.2％(46/109)、44.3％(47/106),妊娠率分别为29.3％(46/157)、37.6％(41/109)、41.5％(44/106),异位妊娠率分别为1.9％(3/157)、1.8％(2/109)、0.9％(1/106),流产率分别为18.6％(8/43)、12.8％(5/39)、20.9％(9/43),活产率分别为22.3％(35/157)、31.2％(34/109)、32.1％(34/106),各指标IVM组分别与对照组1、2比较,差异均无统计学意义(P＞0.05);3组新生儿中,早产率、低出生体质量儿率、新生儿体质量、男女性别比例分别比较,差异均无统计学意义(P＞0.05);IVM组中位促排卵天数6d、中位促性腺激素用量675 IU、治疗总费用(8392±1328)元,均少于对照组1、2,差异均有统计学意义(P均＜0.01);IVM组多胎率4.7％(2/43)和卵巢过度刺激综合征(OHSS)发生率(0)均低于对照组1、2,差异均有统计学意义(P均＜0.01).结论 IVM适用于PCOS不孕症患者,能获得与常规IVF或ICSI相似的妊娠结局,并能减少治疗费用、降低多胎妊娠和OHSS发生率.     

未经促排卵的未成熟卵母细胞行体外成熟治疗不孕症的临床研究

目的探讨未经任何药物刺激的未成熟卵母细胞行体外成熟（IVM）治疗不孕症的临床价值。方法40例不孕患者接受54个IVM周期,其中多囊卵巢综合征（PCOS）不孕患者26例,经其他辅助生育技术失败14例。在未采用任何药物刺激的前提下,于月经周期的第9—12天,在超声引导下经阴道对两侧卵巢内直径≤10mm的卵泡进行穿刺取卵。对取出卵母细胞于体外培养24～48h,待第一极体出现后,进行卵母细胞质内单精于注射（ICSI）,18h后观察受精情况,继续培养24—48h,直至胚胎移植,移植前行激光辅助胚胎孵化。结果54个IVM周期中,有7个周期取消,取消率为13％;共移植周期47个,共获得未成熟卵母细胞857个,平均每周期18．2个。体外培养48h后,卵母细胞成熟率为73．7％（632／857）,正常受精率为75．3％（476／632）,卵裂率为91．2％（434／476）。移植日子宫内膜厚度平均为8．9mm,平均移植胚胎4．3个（2—6个）;1例生化妊娠,19例临床妊娠,每取卵周期的临床妊娠率为35％（19／54）,每移植周期的临床妊娠率为40％（19／47）。26例PCOS不孕患者共移植周期34个,1例生化妊娠,15例临床妊娠,每移植周期的临床妊娠率为44％（15／34）。结论未经促排卵药物刺激的卵母细胞行IVM用于治疗各种原因的不孕症,尤其是PCOS不孕患者,是一种有效的治疗方法。     

未成熟卵母细胞体外培养技术助孕治疗多囊卵巢综合征疗效研究

目的 探讨对多囊卵巢综合征（PCOS）患者行未成熟卵母细胞体外培养技术（IVM）助孕治疗的有效方案。方法 对2010年1月至2012年12月在中信湘雅生殖与遗传专科医院接受体外受精（IVF）助孕治疗的PCOS患者进行回顾性分析。按助孕方案分3组：A组：自然周期IVM（140例）;B组：促性腺激素释放激素激动剂（GnRH-a）+短暂重组卵泡刺激素（r-FSH）刺激再行IVM(152例);C组：长方案降调节后常规体外受精-胚胎移植(505例)。分别比较3种助孕方案对PCOS患者的疗效。结果 （1）临床妊娠率：B组52.59％,C组58.35％,差异无统计学意义（P>0.05）,但均明显高于A组（32.23％）（P<0.05）。（2）人绒毛膜促性腺激素（HCG）日内膜厚度：A组（5.99±1.96）mm,B组（7.87±2.14）mm,C组（11.81±2.20）mm,3组比较差异均有统计学意义（P<0.05）。（3）优胚率：A组（47.23±29.17）%,B组（59.95±30.43）%,C组（65.66±23.35）%,3组比较差异均有统计学意义（P<0.05）。（4）促性腺激素（Gn）用量：A组未使用Gn,B组（600.08±291.21）U,C组（1776.36±693.76）U,3组比较差异均有统计学意义（P<0.05）。（5）并发症发生率：A、B组无卵巢过度刺激综合征（OHSS）发生,C组重度OHSS发生率为2.18％。结论 IVM方案可以有效避免PCOS患者促排卵时OHSS发生,降调节后短暂Gn刺激再行IVM助孕,可以获得与传统长方案IVF相似的临床妊娠率,是PCOS患者经济、安全有效的助孕方式。     

常规体外受精-胚胎移植周期中改行未成熟卵母细胞体外培养的临床研究

目的 探讨常规体外受精-胚胎移植(IVF-ET)周期中改行未成熟卵母细胞体外培养(IVM)的临床疗效果.方法 回顾性分析2008年1月至2009年6月在温州医学院附属第一医院生殖医学中心行常规IVF治疗,在促排卵5-7 d后,双侧卵泡数目≥20个,或用药8-13 d后,卵泡发育缓慢且两侧卵泡数日≥15个,平均最大卵泡直径≤12 mm的155个周期,根据患者意愿,即刻停药取卵、改行IVM治疗60个周期(A组),继续按照IVF常规治疗95个周期(B组).比较两组促性腺激素(Gn)的用量、周期取消率、平均获卵数、受精率、优质胚胎率、移植周期数、卵巢过度刺激综合征(OHSS)的发生率及妊娠结局.结果 周期移植率A组为92%(55/60),B组为63%(60/95),两组比较,差异有统计学意义(P<0.05).平均Gn用昔、平均获卵数、卵裂率、OHSS发生率A组分别为(1030±468)U、(10±6)个、82.2%(231/281)、0,B组分别为(1544±338)U、(14±4)个、94.0%(502/534)、35%(21/60),两组比较,筹异均有统计学意义(P<0.05);两组的受精率和优质胚胎率比较,差异无统计学意义(P>0.05).A组29例临床妊娠,每移植周期的临床妊娠率为53%(29/55),多胎妊娠发生率为14%(4/29);B组28例临床妊娠,每移植周期的临床妊娠率为47%(28/60),多胎妊娠发牛率为32%(9/28);两组比较,差异无统计学意义(P>0.05).结论 常规IVF周期巾改行IVM,不仅可以避免OHSS的发生和减少促排卵药物的使用,还可获得较高的临床妊娠率.     

PCOS患者未成熟卵体外成熟培养后妊娠结局分析

目的:探讨PCOS患者未成熟卵母细胞体外成熟(in vitro maturation,IVM)治疗的有效性和安全性。方法:分析比较PCOS合并不孕症自愿要求行IVM或IVF治疗的患者临床结局。结果:IVF组的平均优质胚胎数明显高于IVM组(P<0.05),两者的临床妊娠率(39.22%vs 40.32%)和着床率(24.37%vs 23.89%)无差异(P>0.05)。IVM组的妊娠丢失率为38.0%,高于IVF患者(21.1%,P<0.05),出生婴儿(单胎)的平均体质量组间无统计学差异(P>0.05)。结论:对于PCOS患者进行IVM治疗可以获得与IVF周期相类似的临床妊娠率和着床率;但是IVM治疗后流产率明显增加。     

未刺激的未成熟卵体外培养治疗多囊卵巢综合征不孕的研究

目的 探讨未刺激的未成熟卵体外培养成熟(IVM)治疗难治性多囊卵巢综合征(PCOS)不孕的疗效.方法 2004-08-2005-08温州医学院附属第一医院生殖中心对56例难治性PCOS不孕患者进行78个周期的IVM治疗.结果 9个周期取消,取消率为11.5%;69个移植周期共获卵1338个,平均每周期19.4个,培养48h后总成熟率为68.5%.正常受精率为70.2%,卵裂率为91.1%.移植目的子宫内膜厚度平均8.7mm,平均移植3.7个胚胎.1例异位妊娠,2例生化妊娠,28例获临床妊娠.每穿刺周期和移植周期的临床妊娠率分别为37.3%(28/75)和40.6%(28/69).28例临床妊娠中,3例孕8～12周流产,1例孕22周因胎膜早破流产,15例已分娩,2例双胎和7例单胎在妊娠中.多胎妊娠率为39.3%(11/28).结论 未经刺激的IVM技术用于治疗难治性PCOS不孕是一种有效的方法,具有简单、经济、最低损伤且可获得与常规体外受精-胚胎移植(ⅣF-ET)相近的临床妊娠率,但多胎妊娠的发生率仍较高.     

Risks of fertilization in vitro

The authors report the results of four IVF attempts, performed on the same couple, in 16 months, because of tubal problems. June 1985 (T1): spontaneous miscarriage after 8 weeks of amenorrhea; october 1985 (T2): negative tap; january 1986 (T3): extra-uterine pregnancy; october 1986 (T4): rejected because of "poor response" and spontaneous extra-uterine pregnancy during the same cycle.     

In vivo development of in vitro fertilized bovine oocytes matured in vivo versus in vitro

In vivo developmental potentials of in vivo and in vitro matured oocytes fertilized in vitro were assessed in cattle. One-cell stages produced from in vivo matured oocytes developed into a pregnancy when transferred to the ampulla part of oviducts of synchronized heifers. In vitro matured oocytes achieved high penetration and cleavage rates but did not develop into pregnancies when transferred to synchronized heifers.     

Oocyte maturation in vitro

In contrast to conventional assisted reproduction techniques in which metaphase II oocytes are retrieved for fertilization in vitro, during the in-vitro maturation of oocytes, immature germinal vesicle stage oocytes are retrieved and matured in the laboratory before fertilization and embryo transfer. Whereas in-vitro maturation is technically more demanding than in-vitro fertilization it carries many potential advantages in terms of lower treatment costs and greater safety, as a result of reducing both the number of clinical consultations and the amount of pharmacological intervention. Although the technology is still experimental, in-vitro maturation has been successfully used for the treatment of patients with polycystic ovarian syndrome, and a number of pregnancies and live births have been reported. Despite these successes and much continuing research effort, the overall efficiency of in-vitro maturation remains low, and neither clinical nor laboratory procedures can be considered to be as robust and routinely feasible as conventional in-vitro fertilization techniques. The immediate goals for in-vitro maturation programmes are therefore to increase germinal vesicle oocyte recovery rates, optimize culture conditions, improve oocyte maturation potential, and after fertilization to produce pregnancy rates that are at least equivalent to those obtained using conventional in-vitro fertilization. This review focuses on the physiology and application of human oocyte maturation in vitro, and will detail the recent advances reported in this rapidly advancing field.     

La capacitation in vitro

Capacitation is a prerequisite for mammalian spermatozoa to fertilize oocytes. Lipids play a crucial role in the structural and functional organization of sperm plasma membrane. Lipid and membrane protein ordering changes dramatically during sperm capacitation but the resulting effects differ according to the regions of the sperm head. Lipids modifications are mainly characterized by a cholesterol efflux, dynamic cholesterol redistribution in particular in the apical zone of the head and also a phospholipids reorganization resulting to the scramblase activation. The existence of lipids ordered microdomains (lipid rafts) has been recently observed in sperm membranes. These lipid and membrane protein movements are believed to play a role in modulating signaling pathways mainly, the AMPc/PKA and ERK pathways. One of the early key events is the activation of adenylate cyclase by high levels of bicarbonate. All these pathways lead finally to the phosphorylation of Tyr-proteins. But capacitation seems to be more complex with the contribution of other kinases (from the PI3K/Akt pathway and phosphotyrosine kinases) towards the phosphorylation of other Ser/Thr and Tyr proteins. The reactive oxygen species (ROS) seem to be important in the control of mechanisms involved in capacitation.     

Follicular development in vitro

There has been tremendous interest in recent years in the culture of oocytes and follicles. Although much of the research using follicle culture aims to increase understanding of the regulation of follicle development, an important goal has been to develop a method that will eventually allow maturation of human oocytes from the primordial follicle to the mature Graafian stage. We are still some way from this at present, although it has now been achieved in the mouse. In this article, we consider various methods of follicle culture for primordial, preantral, and antral follicles. In vitro development of primordial follicles has used primarily whole ovaries or ovarian fragments as a source of follicles. Culture of later stages of follicle development uses mainly isolated follicular units, either whole (with an intact basement membrane and, in some cases, attached thecal cells) or nonintact (oocyte-somatic cell complexes, which may or may not have remnants of basement membranes and/or thecal cells attached).     

A comparison of in vitro maturation and in vitro fertilization for women with polycystic ovaries

OBJECTIVE: To establish the relative success of treatment by unstimulated in vitro maturation (IVM) of oocytes or stimulated in vitro fertilization (IVF) in women with polycystic ovaries undergoing assisted conception treatment. METHODS: The case-control study included 107 IVM and 107 IVF cycles matched for age and cause of infertility. In vitro maturation patients underwent transvaginal recovery of immature oocytes during an unstimulated cycle, in vitro oocyte maturation, and fertilization. Those in the IVF group underwent ovarian stimulation after pituitary suppression. Embryos were transferred in the same cycle in both groups. Main outcome measures included numbers of mature oocytes and embryos produced, and rates of implantation, pregnancy, live birth, and complications. RESULTS: In the IVM group after in vitro culture, 7.8 mature oocytes and 6.1 embryos were obtained per retrieval. With IVF, 12.0 mature oocytes (P <.01) and 9.3 embryos (P <.01) were obtained. The IVM pregnancy and live birth rates per retrieval were 26.2% and 15.9% compared with 38.3% and 26.2% for IVF (nonsignificant). The implantation rate of IVF-derived embryos was higher (17.1% versus 9.5%) than that for IVM (P <.01). There were 12 cases (11.2%) of moderate or severe ovarian hyperstimulation syndrome in IVF patients, compared with none in the IVM group (P <.01). CONCLUSION: Our results suggest that for women with polycystic ovaries who require assisted conception, IVM is a promising alternative to conventional IVF treatment.