未成熟卵母细胞体外成熟培养研究进展

自Pincus发现卵母细胞可在体外成熟至今,未成熟卵母细胞的体外成熟培养一直是国内外生殖医学领域研究者关注的热点.成功的培养未成熟卵母细胞,不仅可避免目前药物诱导排卵引起的多种弊端,还为卵子库的建立展示了广阔的前景.就卵母细胞成熟的调节机理、未成熟卵的获取、体外培养条件及影响体外培养的因素等方面作综述.     

未成熟卵母细胞体外成熟培养研究进展

自Pincus发现卵母细胞可在体外成熟至今,未成熟卵母细胞的体外成熟培养一直是国内外生殖医学领域研究者关注的热点 成功的培养未成熟卵母细胞,不仅可避免目前药物诱导排卵引起的多种弊端,还为卵子库的建立展示了广阔的前景。就卵母细胞成熟的调节机理、未成熟卵的获取、体外培养条件及影响体外培养的因素等方面作综述     

卵母细胞体外成熟机制及分子遗传学研究进展

人未成熟卵母细胞体外培养成熟(in vitro maturation,IVM)是现代生殖医学的研究热点,是当今国际辅助生殖前沿技术.卵母细胞体外成熟是一个非常复杂的过程,不仅包括细胞核的成熟,也包括细胞质成熟;细胞核、质成熟的同步性至关重要,卵母细胞的发育潜能往往因二者的不同步而受阻.就IVM的临床意义、卵母细胞成熟机制、卵母细胞体外成熟非整倍体发生机理做简要综述.     

人未成熟卵母细胞体外成熟影响因素的探讨

随着阴道B超、胞浆内单精子注射(ICSI)技术的开展,未成熟卵母细胞体外成熟培养(IVM)已成为治疗不孕症的一种经济有效的新途径.向IVM培养系统添加促性腺激素、卵泡液、表皮生长因子、血管内皮生长因子、抑制素/激活索、人绒毛膜促性腺激素以及共培养系统有助于卵母细胞体外成熟.根据卵母细胞所处发育阶段选择不同的培养系统,探索卵母细胞的生长和成熟是未来的重要研究方向.     

卵母细胞体外成熟调节的研究进展

虽然已有一定数量的未成熟人卵母细胞体外成熟(IVM)、体外受精-胚胎移植(IVF-ET)后得到的正常后代,但其发育潜能显然不如体内成熟的卵母细胞好,这主要是由于对灵长类动物卵母细胞胞质成熟及卵母细胞获得发育能力方面的分子机理和生理生化过程知之甚少,现有的IVM条件不足以维持卵母细胞的胞质成熟.所以,改良IVM培养基的有效成分,积极探索卵母细胞IVM的条件,成功建立IVM、IVF和体外胚胎培养(IVC)程序,对生殖生物学及生殖工程的发展具有重要意义.就未成熟卵母细胞的IVM调节因素的研究进展进行综述.     

卵母细胞体外成熟机制及分子遗传学研究进展

人未成熟卵母细胞体外培养成熟(invitromaturation,IVM)是现代生殖医学的研究热点,是当今国际辅助生殖前沿技术。卵母细胞体外成熟是一个非常复杂的过程,不仅包括细胞核的成熟,也包括细胞质成熟;细胞核、质成熟的同步性至关重要,卵母细胞的发育潜能往往因二者的不同步而受阻。就IVM的临床意义、卵母细胞成熟机制、卵母细胞体外成熟非整倍体发生机理做简要综述。     

未成熟卵体外培养研究综合进展

尽管体外培养成熟(IVM)技术已基本接近成熟,但其成熟率还很低而且还存在许多不足和缺点.近年来随着医学细胞与分子生物学技术的迅速发展,未成熟卵体外培养研究取得巨大进展,并进行了相关改进.对近几年一些较新的研究发现进行总结,主要从卵母细胞成熟动力学,内分泌学,培养液及培养方案的选择,卵母细胞成熟指标等方面予以综述.     

卵母细胞体外成熟机制及分子遗传学研究进展

人未成熟卵母细胞体外培养成熟（in vitro maturation，IVM）是现代生殖医学的研究热点．是当今国际辅助生殖前沿技术。卵母细胞体外成熟是一个非常复杂的过程，不仅包括细胞核的成熟，也包括细胞质成熟；细胞核、质成熟的同步性至关重要，卵母细胞的发育潜能往往因二者的不同步而受阻。就IVM的临床意义、卵母细胞成熟机制、卵母细胞体外成熟非整倍体发生机理做简要综述。     

小鼠巨细胞病毒对体外成熟卵母细胞超微结构的影响

目的:探讨小鼠巨细胞病毒对小鼠卵母细胞体外成熟过程中超微结构的影响。方法:将小鼠未成熟卵母细胞置于含不同浓度小鼠巨细胞病毒的培养液中培养,观察卵母细胞的生发泡破裂率和第一极体释放率,用共聚焦显微镜和电子显微镜观察体外成熟过程中卵母细胞超微结构的改变。结果:各组卵母细胞生发泡破裂率和第一极体释放率差异无显著性(P>0.05),100TCID50组卵母细胞的超微结构及纺锤体结构未见明显异常。结论:小鼠巨细胞病毒对未成熟卵母细胞体外成熟过程中超微结构无明显不良影响。     

人未成熟卵母细胞体外成熟影响因素的探讨

随着阴道B超、胞浆内单精子注射(ICSI)技术的开展,未成熟卵母细胞体外成熟培养(IVM)已成为治疗不孕症的一种经济有效的新途径。向IVM培养系统添加促性腺激素、卵泡液、表皮生长因子、血管内皮生长因子、抑制素/激活素、人绒毛膜促性腺激素以及共培养系统有助于卵母细胞体外成熟。根据卵母细胞所处发育阶段选择不同的培养系统,探索卵母细胞的生长和成熟是未来的重要研究方向。     

卵母细胞体外成熟技术的临床应用进展

卵母细胞体外成熟(in vitro maturation,IVM)是将处于不同时期的未成熟卵母细胞体外培养成熟的技术,临床不同来源的未成熟卵母细胞在胚胎发育潜能、妊娠率和活产率等方面存在差异。IVM作为一种有效治疗手段可应用于治疗多囊卵巢综合征、卵巢高反应及低反应患者、癌症患者生育力保存等,该技术已在全世界应用并有数千名健康婴儿出生。临床IVM技术的改进主要包括IVM培养基的改良、培养环境及操作流程的优化。目前,随着体外受精(IVF)效率的提高和培养体系的改进,自然周期或温和刺激可能更适合于进行IVF治疗的女性。新的治疗方案提出将自然周期/温和刺激IVF与IVM结合,温和刺激IVF联合IVM治疗方案不仅有望成为目前标准治疗的可行替代方案,而且可能是潜在的一线治疗选择。     

未成熟卵体外成熟技术在卵巢高反应患者IVF-ET中的应用

目的:探讨未成熟卵体外成熟(IVM)技术在卵巢高反应患者体外受精-胚胎移植(IVF-ET)中的应用价值。方法:在IVF-ET促排治疗中,对双卵巢卵泡数过多,有可能发生卵巢过度刺激综合征(OHSS)或继续治疗可能发生重度OHSS的患者,根据其意愿即刻停药,全部取卵改行IVM治疗12个周期(A组)或取部分小卵泡改行IVM治疗,同时保留部分卵泡继续行IVF-ET常规治疗18个周期(B组)。小卵泡体外培养成熟后,通过卵胞浆内单精子注射(ICSI)获得受精卵并行胚胎移植或冷冻。统计分析未成熟卵的成熟率、卵母细胞的受精率、胚胎的发育情况及临床结局。结果:两组30个取卵周期,共获未成熟卵240个,经IVM、ICSI和体外培养后,成熟率、受精率、正常卵裂率及优质胚胎率分别为61.25%(147/240),77.55%(114/147),92.98%(106/114)和29.25%(31/106)。A组8例行IVM新鲜胚胎移植(8周期)4例临床妊娠,A、B两组有8例行IVM解冻胚胎移植(9周期)3例临床妊娠,已有3例分娩。A组12例无OHSS发生,促性腺激素用量少于B组,B组18例中3例有OHSS风险而取消胚胎移植。结论:对常规IVF促排周期中卵巢高反应患者及时改行IVM,可以避免周期取消及OHSS的发生,减少促排卵药物的使用量,同时获得较好的临床妊娠率。     

SCHEDULE TO INJECT IN VITRO MATURED OOCYTES MAY INCREASE PREGNANCY AFTER INTRACYTOPLASMIC SPERM INJECTION

To ascertain the value of using immature oocytes in an intracytoplasmic sperm injection (ICSI) program, the authors designed a schedule, at 5 p.m. on day 1 (the day of oocyte retrieval) and at 8 a.m. and 2 p.m. on day 2, to recognize and inject the in vitro matured (IVM) oocytes. For the 1166 oocytes retrieved in 107 ICSI cycles, 128 (11.0%) were at the stage of metaphase I (MI) and 113 (9.7%) at germinal vesicle. Routine ICSI for metaphase II oocytes was performed at 2 p.m. on day 1 (initial ICSI). In culture medium of human tubal fluid with 15% maternal serum, 85.1% (205/241) immature oocytes progressed to maturation in which 16.4% (21/128) of MI oocytes matured at 5 p.m. of day 1. The rate of normal fertilization for IVM oocytes (58.5%) was not significantly different from that of initial ICSI (64.0%). One patient received a transfer of two fertilized IVM oocytes alone that were injected at 5 p.m. of day 1, maturing from the MI stage, and achieved a normal pregnancy. The fertilized IVM oocytes were replaced along with the embryos from initial ICSI for 40 cycles that led to 14 (35%) clinical pregnancies. In 43 fertilized IVM oocytes donated for research, we observed that cleavage (95.3%) to the 2- to 4-cell stage was not distinct from that of initial ICSI (94.6%); however, the percentage of embryos of grade I and II morphology was significantly smaller (24.4% vs. 62.5%). Only five (11.6%) developed to blastocysts in vitro. Twenty-one fertilized IVM oocytes were frozen for future transfer. A schedule to inject IVM oocytes in ICSI cycles may generate more accessible embryos for fresh transfer or cryopreservation to increase the chance of pregnancy, although the embryo quality was relatively poor.     

Synergistic property of piperonyl butoxide,diethyl maleate,triphenyl phosphate and verapamil hydrochloride with deltamethrin and ivermectin against Rhipicephalus microplus ticks

The present study was taken up to evaluate the synergistic properties of piperonyl butoxide (PBO), diethyl maleate (DEM), triphenyl phosphate (TPP) and verapamil (VER) with deltamethrin (DLM) and ivermectin (IVM) against DLM and IVM resistant tick populations collected from Madhya Pradesh and Punjab states of India. The collected field tick populations were resistant to DLM (Resistance Factor [RF] in the range of 21.71–32.98) and IVM (RF in the range of 1.89–4.98). A strong synergism between DLM and, IVM with PBO and IVM with VER was noticed. The synergistic efficacy of PBO and VER with IVM in reducing the lethal concentration 50 (LC50) value (1.69–5.72 times for PBO and 3.00–10.62 times for VER) of IVM in resistant ticks suggest that a combination of these synergists with IVM can significantly enhance the effectiveness of IVM against IVM-resistant Rhipicephlaus microplus populations gradually establishing in the different parts of the country. The synergistic efficiency of PBO with DLM in reducing the LC50 value was 2.65 and 18.01 times, respectively, against DLM- resistant two R. microplus populations (KTN and LDH). The study revealed the gradual establishment of DLM and IVM resistant populations in the surveyed states suggesting the need to adopt required resistance management strategies. The use of synergists with DLM and IVM has emerged as an effective approach for controlling the acaricide-resistant ticks.     

Improvement of embryonic development and clinical outcomes of germinal vesicle stage oocytes using a microvibration culture system

ABSTRACT

In vitro maturation (IVM) has evolved as a clinical treatment option in assisted reproductive technology. However, the poor developmental potential of germinal vesicle (GV)-stage oocytes is still suboptimal. This study’s objective was to evaluate the effect of a microvibration culture system (MVC) during IVM and/or in vitro culture (IVC) on the clinical outcomes and the embryonic development potential of human GV-stage oocytes collected from human chorionic gonadotropin (HCG)-primed IVM and fertilization-embryo transfer (IVM/F-ET) cycles of patients with polycystic ovaries (PCO). A total of 206 HCG-primed IVM/F-ET cycles were divided into four groups according to the microvibration and static culture system applied during IVM and/or IVC: Group SS (static system during both IVM and IVC); Group SV (static system during IVM alternated with microvibration system during IVC); Group VS (microvibration system during IVM alternated with static system during IVC), and Group VV (microvibration system during both IVM and IVC). The results indicate that the rates of in vitro MII oocytes per cycle, fertilization, and cleavage were not significantly different between the groups. The rate of good-quality embryos in Group SV tended to be higher than the rate in Groups SS and VS, but there was no significant difference between Group SS and Group SV. Clinical pregnancy, implantation, and live birth rates of Groups SV and VS were slightly higher than those of Group SS. However, the rate of good-quality embryos with at least six cells on day 4, the clinical pregnancy, implantation, and live births in Group VV were significantly higher than those in Group SS. These results indicate that, compared with the static culture system, the MVC system applied for both IVM and IVC seems to improve the clinical outcomes and the quality of embryos of GV oocytes derived from HCG-primed IVM/F-ET cycles in PCO patients.

Abbreviations: PCO: polycystic ovaries; HCG: human chorionic gonadotropin; GV: germinal vesicle; MII: metaphase II; IVM: in vitro maturation; IVF: in vitro fertilization; IVC: in vitro culture: MVC: microvibration culture; SC: static culture; ICSI: intracytoplasmic sperm injection; IVM/F-ET: IVM and fertilization-embryo transfer; AMH: anti-Mullerian hormone; OHSS: ovarian hyperstimulation syndrome     

Efficacy and safety of drug combinations in the treatment of schistosomiasis, soil-transmitted helminthiasis, lymphatic filariasis and onchocerciasis

This review concerns the efficacy and safety of combinations of various drugs, including albendazole (ALB), diethylcarbamazine (DEC), ivermectin (IVM), mebendazole and praziquantel (PZQ). There were no significant pharmacokinetic interactions when ALB-PZQ, ALB-DEC, ALB-IVM or ALB-IVM-PZQ were co-administered. ALB did not add to the cure rate of PZQ in the treatment of Schistosoma japonicum, S. mansoni and S. haematobium. ALB and DEC in combination and alone were ineffective against S. haematobium infections. No combinations (ALB-PZQ, ALB-IVM and ALB-DEC) were superior to ALB against Ascaris lumbricoides and hookworm infections, whilst IVM, but not PZQ or DEC, added to the effect of ALB in the treatment of Trichuris trichiura. Results with ALB added to single-drug therapy with IVM or DEC against lymphatic filariasis were inconclusive, but DEC and IVM in combination appeared to be superior to DEC or IVM alone. None of the drug combinations against lymphatic filariasis showed more adverse reactions than single-drug therapy. In onchocerciasis patients, ALB and IVM were safe in those also infected with lymphatic filariasis, but were not superior to IVM alone. Existing policies are based on limited knowledge. Well conducted, comparative, randomised controlled studies would greatly aid in the future use of these drug combinations.     

Cysteamine supplementation of in vitro maturation medium, in vitro culture medium or both media promotes in vitro development of buffalo (Bubalus bubalis) embryos

The effects of supplementation of in vitro maturation (IVM) or in vitro culture (IVC) or both IVM and IVC media with cysteamine on the yield, hatching rate (HR) and total cell number (TCN) of buffalo blastocysts were examined. Oocytes obtained from slaughterhouse buffalo ovaries were subjected to IVM and IVF. The IVM or IVC media were supplemented with 0, 50, 100 or 200 microm cysteamine. Supplementation of IVM medium with 50 microm cysteamine increased (P < 0.01) the cleavage rate and blastocyst yield without affecting the HR and TCN whereas a higher concentration of 200 microm significantly (P < 0.05) reduced the blastocyst yield but not TCN. Similar increases in blastocyst yield, without any effect on HR and TCN were observed after supplementation of the IVC medium with 100 (P < 0.01) or 50 microm (P < 0.05) cysteamine, whereas 200 microm cysteamine was ineffective. Supplementation of both IVM medium with 50 microm cysteamine and of IVC medium with 100 microm cysteamine increased the yield of blastocysts and hatched blastocyst by over 100% (P < 0.01) compared with the controls without any adverse effects on HR or TCN. The results of the present study suggest that supplementation of both IVM and IVC media improves the yield of blastocysts without compromising their health.     

Research Progress of In vitro Oocyte Maturation

In vitro maturation (IVM) has been used in clinical settings for 30 years. The merits of IVM include that it needs a relatively small amount of hormones and sho...     

二甲双胍治疗对多囊卵巢综合征患者未成熟卵体外培养结局的影响

目的:评估二甲双胍治疗对多囊卵巢综合征(PCOS)患者未成熟卵母细胞体外培养成熟(IVM)相关结局的影响。方法:选择2005年8月～2007年8月在该中心行IVM治疗的难治性PCOS不孕患者75例共106个周期,根据有无加用二甲双胍治疗及治疗的不同方案分为治疗A组(16例共20个周期)、治疗B组(15例共23个周期)及未治疗C组(44例共63个周期),比较二甲双胍治疗对IVM相关结局的影响。结果:三组的获卵数、受精率、卵裂率、种植率、穿刺周期的临床妊娠率及移植周期的临床妊娠率等指标差异均无统计学意义(P0.05);A、B两组的卵母细胞体外成熟率、优质胚胎数及每周期移植胚胎数差异无统计学意义,但显著高于C组,差异有统计学意义(P0.05);A、B、C三组流产率分别为0.00%、14.30%及20.00%,A、C两组间差异有统计学意义(P0.05)。结论:对于难治性PCOS患者,IVM技术可获得较高妊娠率,而二甲双胍治疗可在一定程度上改善IVM结局。