In-vitro maturation of human oocytes: before or after vitrification?

Purpose

This study aims to determine if in-vitro maturation (IVM) of human immature oocytes should be performed before or after vitrification.

Methods

A total of 184 immature oocytes were randomly divided into two different groups: 100 were vitrified at metaphase II (MII) stage 24 h-48 h after IVM (group 1) and 84 were immediately vitrified at germinal vesicle (GV) or metaphase I (MI) stages and in vitro matured after warming (group 2).

Results

Survival rate after warming was similar in both groups (86.9% versus 84.5%). However, oocyte maturation rate per collected oocyte was significantly higher for oocytes matured before vitrification (group 1, 46%) than for oocytes vitrified before IVM (group 2, 23.8%) (p < 0.01). Consequently, the number of MII oocytes inseminated per oocyte collected was significantly higher for group 1 (40%) than for group 2 (23.8%) (p < 0.05).

Conclusion

IVM procedure is more efficient when it is performed before oocyte vitrification.     

多囊卵巢综合征患者中未成熟卵母细胞体外培养成熟后体外受精的初步研究

目的:探讨未成熟卵母细胞体外培养成熟后体外受精、胚胎培养技术在多囊卵巢综合征患者中的初步应用及其影响因素。方法:用小剂量促性腺激素促使卵泡生长后,根据优势卵泡直径分为2组,直径6～8mm者为组1,10—12mm者为组2。采集未成熟卵母细胞,经体外培养成熟后,再进行体外授精和胚胎培养。结果:组1的GV期卵的成熟率和受精率低于组2者,但两组的MI期卵的成熟率、受精率没有明显差别。两组卵裂率没有明显差别,但形成胚胎的质量组2优于组1。总计成熟率69.3％,成熟卵中正常受精率61.5％。结论:可以用小荆量Gn促使卵泡生长后,采集未成熟卵,用体外成熟-体外授精(IVM/IVF)的方法使多囊卵巢患者在避免OHSS的情况下获得质量良好的胚胎。     

超排卵周期未成熟卵体外培养的研究

目的:研究来源于超排卵周期中的未成熟卵在拆除卵丘细胞后进行体外成熟培养(IVM)的成熟、受精及胚胎发育能力,探讨IVM技术的临床应用。方法:选取46名体外受精/卵胞浆内单精子显微注射-胚胎移植(IVF/ICSI-ET)患者为研究对象,比较MI和GV期不成熟卵的体外成熟情况,并比较体内成熟卵和体外成熟卵进行ICSI后的正常受精、异常受精、卵裂和优质胚胎形成情况。结果:体外培养中69.8%的MI期卵和77.2%的GV期卵均在24小时内达到成熟,其24小时和48小时的成熟率、总成熟率均无明显差异(P>0.05)。体外成熟卵与体内成熟卵相比较,正常受精率、异常受精率和卵裂率均无明显差异(P>0.05),优质胚胎形成率较低,差异有显著性(P<0.05)。结论:常规超排卵周期中的未成熟卵在拆除卵丘细胞后能够继续体外发育成熟,具有与体内成熟卵相似的ICSI受精、卵裂能力。虽然优质胚胎的形成率低于体内成熟卵,但增加了可移植胚胎和冷冻胚胎数量,提高了助孕成功率。     

Vitrification of human immature oocytes before and after in vitro maturation: a review

The use of immature oocytes subjected to in vitro maturation (IVM) opens interesting perspectives for fertility preservation where ovarian reserves are damaged by pathologies or therapies, as in PCO/PCOS and cancer patients. Human oocyte cryopreservation may offer some advantages compared to embryo freezing, such as fertility preservation in women at risk of losing fertility due to oncological treatment or chronic disease, egg donation and postponing childbirth. It also eliminates religious and/or other ethical, legal, and moral concerns of embryo freezing. In addition, a successful oocyte cryopreservation program could eliminate the need for donor and recipient menstrual cycle synchronization. Recent advances in vitrification technology have markedly improved the oocyte survival rate after warming, with fertilization and implantation rates comparable with those of fresh oocytes. Healthy live births can be achieved from the combination of IVM and vitrification, even if vitrification of in vivo matured oocytes is still more effective. Recently, attention is given to highlight whether vitrification procedures are more successful when performed before or after IVM, on immature GV-stage oocytes, or on in vitro matured MII-stage oocytes. In this review, we emphasize that, even if there are no differences in survival rates between oocytes vitrified prior to or post-IVM, reduced maturation rates of immature oocytes vitrified prior to IVM can be, at least in part, explained by underlying ultrastructural and biomolecular alterations.     

Cryopreservation of human germinal vesicle stage and in vitro matured M II oocytes: influence of cryopreservation media on the survival, fertilization, and early cleavage divisions

OBJECTIVE: To study the influence of low-sodium cryopreservation media (CPM) on the survival and development of frozen-thawed germinal vesicle (GV) stage and in vitro matured human oocytes. DESIGN: Prospective experimental study. SETTING: Academic hospital-based fertility center. PATIENT(S): Experimental groups: Oocytes cryopreserved at the GV (group A, n = 63 and group B, n = 64) or M II stage (group C, n = 62) with use of conventional (group A) or low-sodium CPM (groups B and C). Control groups: Sibling GV stage oocytes subjected to in vitro maturation (IVM; control group A, n = 64; control group B, n = 64).Intervention(s): IVM, intracytoplasmic sperm injection and subsequent culture. MAIN OUTCOME MEASURE(S): Rates of survival, maturation, fertilization, and cleavage. RESULT(S): The postthaw survival was significantly lower in groups A (57.1%) and B (48.4%) compared to C (84.4%). In group A, maturation and cleavage rates were significantly lower, and fertilization rate was similar to controls (GVBD: 72.2% vs. 90.6%; progression to M II: 33.3% vs. 76.6%; cleavage: 42.9% vs. 88.2%; and fertilization: 58.3% vs. 69.4% in group A vs. control group A, respectively). There was no such difference in group B. In group C, despite a slight but significant lowering of the rate of 2 PN and an increase in that of 3 PN (2 PN: 47.4% vs. 70.2% and 3 PN: 15.8% vs. 3.2% in group C vs. total controls, respectively), embryonic cleavage per GV oocyte was significantly higher (25.8%) compared to group A (4.8%) but not to group B (15.6%). The rate of maturation and cleavage per surviving GV oocyte was significantly higher in group B than group A. CONCLUSION(S): Low-sodium-based CPM is beneficial for in vitro matured M II stage oocytes and is significantly better than the conventional sodium-based media for the GV stage oocytes.     

The effect of vitrification on ultrastructure of human in vitro matured germinal vesicle oocytes

Objective

To describe the possible effects of cryotop vitrification on maturation rate and ultrastructural morphology of human in vitro matured germinal vesicle (GV) oocytes.

Study design

A total of 301surplus immature GV oocytes obtained from infertile patients were allocated into two groups: (i) GV oocytes (n = 150) matured in vitro (fIVM), and (ii) GV oocytes (n = 151) that were first vitrified, then matured in vitro (vIVM). Supernumerary fresh in vivo matured oocytes (n = 10) were used as controls. The maturation media was Ham's F10 supplemented with FSH + LH and human follicular fluid. After 36 h of incubation, the oocytes were investigated for nuclear maturation and ultrastructural changes using transmission electron microscopy (TEM).

Results

Oocyte maturation rates were reduced (P < 0.001) in vIVM (45.92%) in comparison with fIVM oocytes (75.33%). The rate of degeneration was also significantly higher in vIVM than in the fIVM group (44.4% vs. 6.0%). Large and numerous mitochondria and minute vesicles of smooth endoplasmic reticulum (SER) complexes (MV complexes) were observed in both fIVM and vIVM groups. In addition, TEM revealed a drastic reduction in amount of cortical granules (CGs) at the cortex of vitrified-warmed GV oocytes, as well as appearance of vacuoles and small mitochondria-SER aggregates in the ooplasm.

Conclusion

The vitrification procedure is associated with ultrastructural alterations in specific oocyte microdomains, presumably related to the reduced competence of cryopreserved oocytes for maturation. This information emphasizes the need for further work on advancing the cryotechnology of human oocytes.     

无刺激周期未成熟卵母细胞体外培养用于多囊卵巢综合征不孕患者治疗的临床观察

目的探讨无刺激周期未成熟卵母细胞体外培养在治疗多囊卵巢综合征(PCOS)不孕患者的应用价值。方法对70例PCOS患者,在人工周期或自然周期第10～12天行阴道B超监测,如双侧卵巢中无直径>8mm的卵泡,即肌内注射人绒毛膜促性腺激素10000IU,36h后经阴道穿刺取卵,将取出的未成熟卵母细胞进行体外成熟及体外受精胚胎移植。结果共进行体外成熟周期94个,移植周期75个。共获得卵母细胞1283个,获得成熟838个(65.3%,838/1283),其中受精553个(66.0%,553/838),获得胚胎402个(48.0%,402/838),获得优质胚胎199个(23.7%,199/838)。生发泡期卵母细胞的成熟率、受精率、获得胚胎率及优质胚胎率,分别为67.7%、66.4%、47.6%及24.1%;第1次减数分裂中期的卵母细胞,分别为69.7%、71.7%、52.2%及26.1%,生发泡期与第1次减数分裂中期比较,差异均无统计学意义(P>0.05)。无法评价的卵母细胞的成熟率、受精率、获得胚胎率及优质胚胎率,分别为44.8%、53.8%、46.2%及16.9%,与生发泡期及第1次减数分裂中期比较,差异均有统计学意义(P<0.05)。75个移植周期中,18例获得临床妊娠,妊娠率为24%(18/75)。结论无刺激周期未成熟卵母细胞体外成熟及体外受精胚胎移植可用于PCOS不孕患者的治疗,并能取得一定的妊娠成功率。     

Fertility preservation for breast-cancer patients using IVM followed by oocyte or embryo vitrification

Unstimulated in-vitro maturation (IVM) cycles are considered for fertility preservation in breast cancer due to avoidance of ovarian stimulation and shortened time to oocyte retrieval. This study evaluated the efficacy of this approach in a retrospective cohort analysis of 66 patients with breast cancer. Immature oocytes were collected and matured in vitro and then either vitrified or fertilized and preserved as vitrified embryos. In group 1 (vitrified oocytes, n = 35), the average number of oocytes retrieved was 11.4 ± 8.8, the maturation rate was 64.2% and an average of 7.9 ± 6.6 oocytes were vitrified per patient treated. The median duration from the first evaluation to oocyte retrieval was 8 days. In group 2 (vitrified embryos, n = 31) the average number of oocytes retrieved was 9.7 ± 6.4, the maturation rate was 53.2% and an average of 5.8 ± 2.7 mature oocytes were available for fertilization/patient. The fertilization rate was 77.8%, resulting in 4.5 ± 2.7 vitrified embryos/patient. The median duration from the first evaluation to oocyte retrieval was 13 days. Calculated pregnancy rates per vitrified oocyte and embryo were 3.8% and 8.1%, respectively. IVM can be considered a useful option for fertility preservation in breast-cancer patients.Breast cancer represents about 30% of malignant tumours occurring in women of childbearing age. Approximately 10–15% of breast cancers are diagnosed in women of reproductive age. Over the past two decades, earlier diagnosis and highly effective systemic therapies have led to reductions in mortality. However, women undergoing treatment for breast cancer may suffer from fertility problems in the future. A number of methods are currently used to preserve fertility in young breast cancer patients including ovarian freezing, IVF and preservation of oocytes and embryos and the use of drugs to protect the ovary during chemotherapy. A newly described method, in-vitro maturation, was used in our study. Freezing the eggs of breast cancer patients using this technique was found to be a useful treatment option due to two advantages: (i) no hormones were used and (ii) short duration of treatment. Our study evaluates the efficacy of this treatment approach for patients suffering from breast cancer.     

Human oocyte vitrification: in-vivo and in-vitro maturation outcomes

This study aimed to evaluate oocyte vitrification efficiency using in-vivo matured (IVO) versus rescued in-vitro matured (IVM) oocytes. The results show that oocyte survival (85% versus 81%), fertilization (86% versus 76%) and cleavage rate (98% versus 89%) was not significantly different in IVO oocytes compared with rescued IVM sibling oocytes. The fertilized oocytes from IVO and IVM groups were cultured to blastocyst stage; however, embryo development was significantly reduced in the rescued IVM group (72% versus 15%). Embryo transfer was only performed with the embryos derived from IVO oocytes on day 5; 42 blastocysts were transferred to 18 recipients; 16 of 18 recipients had positive beta-human chorionic gonadotrophin and a total of 26 fetal cardiac activities were detected in 15 recipients (implantation: 26/42, 61.9%). Ten of the 15 recipients have delivered 19 healthy babies, and the other five pregnancies are still ongoing. These data indicate that the combination of oocyte vitrification and rescued IVM not only yield a new strategy to extend the pool of total fertilizable oocytes, but also demonstrate that the efficiency of vitrified/warmed oocytes can be comparable to fresh oocytes with regard to clinical outcomes.     

Contribution of rescue in-vitro maturation versus double ovarian stimulation in ovarian stimulation cycles of poor-prognosis women

Research questionMaximizing the number of oocytes and embryos obtained in the shortest possible time is of considerable potential clinical significance for women with poor prognosis. The aim of this study was to develop a clinically applicable strategy to obtain more oocytes and viable embryos in one menstrual cycle for poor-prognosis women.DesignProspective cohort study of 146 women with poor prognosis who received rescue in-vitro maturation (IVM) (n = 50) or double ovarian stimulation (DuoStim) (n = 96) between December 2015 and February 2018. Outcomes, number of oocytes retrieved and matured, and embryo developmental potential were compared between the two groups.ResultsThe rates of mature oocytes, available embryos and top-quality embryos from luteal phase stimulation (LPS) of DuoStim were all significantly higher than those derived from the immature oocytes of rescue IVM (P < 0.05). The relative contributions of LPS in the DuoStim group for proportion of mature oocytes, available embryos and top-quality embryos were all significantly higher than IVM in the rescue IVM group (P < 0.001). The overall cancellation rate of no oocyte or available embryo significantly decreased from 30.21% to 9.38% (P < 0.001) when DuoStim was carried out, which decreased from 24.00% to 12.00% with no significant difference in the rescue IVM group when immature sibling oocytes were matured in vitro.ConclusionRescue IVM and DuoStim can contribute more competent oocytes and viable embryos in the shortest possible time for poor-prognosis women, of which DuoStim may be more efficient.     

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.     

人类未成熟卵母细胞玻璃化冷冻研究

目的:探讨玻璃化冷冻未成熟卵母细胞的有效性。方法:根据有无颗粒细胞将实施玻璃化冷冻的GV期卵母细胞分为含颗粒细胞(非裸卵)组和不含颗粒细胞(裸卵)组;将部分GV期卵母细胞体外培养至MⅡ期卵母细胞实施玻璃化冷冻,比较非冷冻IVM组与MⅡ卵玻璃化冷冻组间、裸卵组与非裸卵组间的存活率、成熟率、受精率、卵裂率及囊胚形成率。结果:非裸卵组的成熟率大于裸卵组(P<0.05),而存活率、受精率、2-细胞形成率、>2-细胞形成率之间均无统计学差异(P>0.05)。另外,非冷冻IVM组与GV玻化组间成熟率、受精率、卵裂率均存在显著性差异(P<0.05);非冷冻IVM组与MⅡ期卵玻化组间成熟率、受精率、卵裂率间均存在统计学差异(P<0.05);GV玻化组与MⅡ玻化组间存活率、成熟率、受精率、卵裂率间均无统计学差异(P>0.05)。结论:玻璃化冷冻未成熟卵母细胞需要保留颗粒细胞,同时初步构建了人GV期卵的玻璃化冷冻联合IVM技术的雏形。     

表皮生长因子卵丘细胞对卵母细胞体外成熟的影响

目的：探讨表皮生长因子（EGF）、卵丘细胞人类生殖泡（germinal vesicle ,GV)期不成熟卵母细胞体外成熟的影响，方法：将EGF和促性激素（gonadotropin,Gn）联合应用于不成熟卵母细胞的体外培养，并同单纯应用Gn相对照观察EGF的影响。同时将卵母细胞分为卵丘卵母细胞复合物（CEO组）及裸卵（DO组）进行培养，了解卵丘细胞在体外培养中的作用。结果：EGF可以提高DO组的MⅡ期率（P＜0．01），对于CEO组应用EGF后可以促进胞质的成熟，提高受精能力（P＜0．05）。关于卵丘细胞的作用，CEO组的MⅡ期率为68．8％，明显高于DO组的46．7％（P＜0．01），CEO组的卵裂率为72．7％，高于DO组的35．7％（P＜0．05）。结论：EGF可以直接促进不成熟裸卵体外成熟，提高其MⅡ期率，与Gn联合应用后可进一步促进卵丘卵母细胞复合物胞质的成熟，提高受精率，保留卵丘细胞利于卵母细胞质的发育，提高不成熟卵母细胞的体外成熟能力，使MⅡ期率，卵裂率升高。     

Correlation between human follicular diameter and oocyte outcomes in an ICSI program

Purpose: To determine the correlation between the follicular sizes and oocyte recovery, metaphase II oocyte recovery, fertilization rate and good embryo quality from mature and immature oocytes in an intracytoplasmic sperm injection (ICSI) program. Methods: 991 follicles obtained from 72 ICSI cycles were classified into three groups according to their diameters as measured by transvaginal ultrasound including group A (<10 mm), group B (10–14 mm), and group C (>14 mm). All obtained oocytes were classified according to their nuclear maturation: germinal vesicle (GV), metaphase I (MI) and metaphase II (MII). Mature oocytes underwent ICSI while immature oocytes were further cultured until maturity before ICSI was performed. The rates of fertilization and good quality embryos at day 3 were evaluated. Results: A progressive and significant increase in the rates of oocyte recovery and MII oocyte recovery were observed from group A follicles compared to the other groups (p < 0.001). The fertilization rate of mature and in vitro matured oocytes, as well as the rate of good quality embryos showed a tendency to increase from group A to group C follicles, but not significantly. The corresponding fertilization rates were 78 and 55.3% (p < 0.001) for mature and in vitro matured oocytes, respectively. Conclusion: Collection of oocytes from small follicles, especially with a mean diameter less than 10 mm, and in vitro maturation of immature oocytes before fertilization may allow the total number of good quality and transferable embryos to be increased.     

The impact of vitrification on murine germinal vesicle oocyte In vitro maturation and aurora kinase A protein expression

Purpose

Investigate the effect of vitrification on in vitro maturation (IVM) and expression of Aurora kinases A, B, and C in germinal vesicle (GV)-stage oocytes.

Methods

GV-stage oocytes from B6D2F1 female mice 7–11 weeks of age were vitrified after collection, thawed, and matured in vitro for 0, 4, 8, and 12 h (hrs). The rate of germinal vesicle breakdown (GVBD), spindle apparatus assembly, and Aurora kinase mRNA and protein expression during IVM was measured.

Results

Oocyte vitrification was associated with significant delays in both GVBD and normal spindle apparatus assembly at 4 and 8 h of IVM (p < 0.05). There was no difference in mRNA levels between control and vitrified oocytes for any of the Aurora kinases. Aurora A protein levels were reduced in vitrified compared to control oocytes at 0 h (p = 0.008), and there was no difference at 4 and 8 h (p = 0.08 and 0.69, respectively) of IVM.

Conclusions

Vitrified oocytes have delayed GVBD and normal spindle assembly during in vitro maturation. Reduced levels of Aurora A protein immediately post-thaw may be associated with the impaired oocyte maturation manifested by the delayed progression through meiosis I and II, and the atypical timing of the formation of meiotic spindles in vitrified GV-stage oocytes.     

Comparison of Quinn's Advantage fertilization medium and tissue culture medium 199 for in vitro maturation of oocytes

ObjectiveThe purpose of the study was to compare the Quinn's Advantage fertilization medium (Q1) and the tissue culture medium 199 (TCM199) for in vitro maturation (IVM) of oocytes and ammonium production during IVM.Materials and methodsThe immature murine oocytes were randomly added into Q1 and TCM199. Ammonium concentrations were measured at the start and after 18 hours of IVM, and the mature oocytes were fertilized and cultured into blastocysts. The blastocysts were then stained for inner cell mass (ICM) and trophectoderm.ResultsThe maturation rate was higher in Q1 than in TCM199 (85.7% vs. 76.6%, p = 0.024). The fertilization and blastocyst rates were slightly higher in Q1, but not significant. Differential staining of the blastocysts showed slightly higher ICM ratio in the blastocysts derived from Q1. Mean ammonium concentrations in Q1 and TCM199 at Time 0 were 184.9 and 339.2 μg/dL, respectively (p = 0.05), and after 18 hours of IVM were 268.7 and 443.6 μg/dL, respectively (p = 0.045). Addition of ammonium chloride into Q1 adversely affects IVM.ConclusionQ1 is superior to TCM199 in terms of oocyte maturation, which may be due to lower ammonium concentration.     

Supplementation with low concentrations of melatonin improves nuclear maturation of human oocytes in vitro

Purpose

Studies in bovine and porcine have indicated that melatonin (MT) could induce meiotic maturation of immature oocytes in vitro. The object of the current study was to investigate if MT could ameliorate human oocytes maturation during rescue in vitro maturation (IVM).

Methods

Two hundred seventy eight germinal vesicle (GV) oocytes and 451 (MI) metaphase I oocytes were vitrified, thawed and then matured in vitro. All the oocytes were randomly allocated into six groups in which the oocytes were cultured in medium supplemented with different concentrations of MT (0, 10⁻², 1, 10², 10⁴, 10⁶ nM) and nuclear maturation was evaluated at 6 h, 12 h, 18 h, 24 h and 48 h of culture.

Results

The optimal MT concentration for both GV and MI oocytes was 1 nM. At 24 h of culture, nuclear maturation rate of MI oocytes cultured in 1 nM MT medium was significantly higher than other groups (P < 0.05); Nuclear maturation rate of GV oocytes cultured in 1 nM MT medium was also significantly higher than the control group (P < 0.05). On the other hand, decreased nuclear maturation rate was observed in the high MT concentration group (10⁶ nM).

Conclusions

The current study demonstrated that low concentration of exogenous MT could ameliorate nuclear maturation of human oocyte during rescue IVM, while high concentration of MT presented negative effects.     

Influence of serum from pregnant women and selected pregnancy hormones on in-vitro maturation of murine oocytes

Maturation medium for in-vitro maturation (IVM) is usually supplemented with serum. The objective of this study was to investigate whether serum from pregnant women had any adverse effects on IVM, and if so, whether the hormonal changes in pregnancy played a role. Serum was obtained from 40 pregnant women and was analysed for oestradiol, prolactin, beta-human chorionic gonadotrophin (beta-HCG), progesterone and testosterone. In the first part of the study, immature murine oocytes were cultured in maturation media supplemented with various sera. The control group was supplemented with 10% serum from fertile women, and groups 2-5 were respectively supplemented with 10, 20, 40 and 80% serum from pregnant women. In the second part of the study, immature murine oocytes were matured in the medium with the addition of different hormones according to the concentrations found in the serum of pregnant women. Compared with the control group, the maturation and fertilization rates were significantly lower in medium supplemented with 10% serum from pregnant women (59.7 versus 71.5% and 60.1 versus 77.0% respectively, both P < 0.05). There was a trend of decreasing maturation and fertilization rates with increasing concentrations of serum from pregnant women. Addition of prolactin, beta-HCG, progesterone and testosterone also impaired oocyte maturation.     

多囊卵巢综合征患者卵丘形态与未成熟卵母细胞发育潜能关系

目的 研究多囊卵巢综合征患者无刺激周期取出的不同形态未成熟卵母细胞的发育潜能。方法 43例PCOS不孕患者进行了47个未成熟卵母细胞体外成熟培养（IVM）周期。所有患者均未经促卵泡素刺激,予以HCG36h后取卵。根据取出的卵-冠-丘复合物形态将其分为3组：卵丘紧密组、卵丘松散组、无卵丘组。比较3组的体外成熟率、受精率和优质胚胎率。结果 47个IVM周期共收集未成熟卵母细胞874枚,体外成熟率61.19%,受精率71.07%,着床率13.13%。卵丘松散组的体外成熟率明显高于卵丘紧密组（72.26%vs49.54%,P〈0.05）,受精率、优质胚胎率三组间无差异。结论 PCOS患者无刺激周期取出的未成熟卵母细胞中,卵丘松散、扩张的卵母细胞具有更好的体外成熟潜力。     

Strategies in human in-vitro maturation and their clinical outcome

The basis of in-vitro maturation (IVM) is the maturing in vitro of oocytes from the germinal vesicle (GV) stage of development to the metaphase II stage. Experience in handling immature oocytes has been obtained from two main groups. The first group is women suffering from polycystic ovarian syndrome, who are extremely sensitive to stimulation with exogenous gonadotrophins in assisted reproduction, and have a significant risk of developing ovarian hyperstimulation syndrome (OHSS). The second group is regular cycling women with normal ovaries referred for IVF due to severe male infertility. In both groups, aspiration of immature oocytes has been performed in unstimulated cycles and after priming with human chorionic gonadotrophin or FSH respectively. Clinical pregnancy rates of 24% per aspiration have been obtained. Children born after IVM appear to be healthy. These data, taken together, suggest that in future, immature oocyte retrieval combined with IVM could replace conventional IVF in selected patients.