Mouse embryo quality control for toxicity determination in the Norfolk in vitro fertilization program

Ongoing quality control is necessary as part of the maintenance and improvement of a successful human in vitro fertilization (IVF) program. Using a mouse quality-control culture system, several instrument preparation protocols were reevaluated to determine their efficiency in the control or elimination of potential toxicity. Dilute concentrations of urine and endometrial fluid were also tested. Medium rinsed through laparoscope and aspiration needles failed to support embryo development. This effect was reversed in needles that were pretreated with rinses of Dulbecco's phosphate-buffered saline. Endometrial fluid demonstrated no obvious toxic effect, but urine-exposed embryos arrested in the two-cell state. The importance of periodic evaluation of materials and their pretreatment before use in in vitro fertilization of human oocytes is essential to ensure control of potentially toxic substances.     

体外受精与胚胎移植前输卵管积水的处理

目的　探讨输卵管积水的处理方法对此后的体外受精与胚胎移植 (IVF-ET)治疗效果的影响。方法　回顾分析 1999年 2月至 2001年 4月期间中山大学附属第一医院生殖医学中心 73例有输卵管积水病史者行IVF-ET治疗的 79个周期的资料。根据在IVF-ET前对患者的输卵管积水的处理方式分A组 27个周期 (25例 ):未经手术治疗行IVF ET;B组 25个周期(23例):已行双侧输卵管切除术;C组 27个周期 (25例 ):已行输卵管积水造口术。D组:以同期因其他输卵管因素行IVF-ET治疗的 913个周期 (875例 )的资料作为对照。结果　B组获卵数最少,较C、D组差异有显著意义(P<0.05);各组的胚胎植入率分别为 9 4%、11.5%、21.2%和 17.4%,A组与C、D组间差异有显著意义(P<0.05)。各组的流产率分别为 50.0%、33.3%、25.0%和 15.3%,A组与D组间的差异有显著意义(P<0.05)。结论　输卵管积水对IVF-ET的胚胎植入构成不良影响,并增加流产的发生,在采用IVF-ET前对输卵管积水进行适当处理,有助于提高胚胎植入率,改善治疗效果。较之输卵管切除术,输卵管造口术对卵巢功能的影响可能更小。     

体外受精与胚胎移植前输卵管积水处理方式的探讨

目的:探讨体外受精与胚胎移植(IVF-ET)前输卵管积水的不同处理方式对IVF-ET结局的影响。方法:回顾分析2002年1月至2004年12月因输卵管因素不孕接受IVF-ET治疗的346个治疗周期的资料。按输卵管的情况分为4组,A组:输卵管阻塞,未发现输卵管积水者,共151个治疗周期;B组:IVF-ET前输卵管积水未行手术治疗者,共73个治疗周期;C组:IVF-ET前输卵管积水行输卵管伞端造口术者,共43个治疗周期;D组:IVF-ET前输卵管积水行输卵管切除术者,共79个治疗周期。结果:各组的临床妊娠率分别为48.34%、21.92%、34.88%、44.30%,异位妊娠率分别为1.37%、12.50%、6.67%、2.86%,种植率分别为20.36%、5.87%、12.28%、19.38%,B组的临床妊娠率及种植率较A组、D组明显降低,B组、C组的异位妊娠率均较A组、D组升高。结论:输卵管积水对IVF-ET的结局有负面影响,IVF-ET前输卵管积水,先行手术治疗,可改善IVF-ET的结局。较之输卵管造口术,输卵管切除术可减少异位妊娠的发生率及输卵管积水的复发率。     

Serum is not necessary in human in vitro fertilization, early embryo culture, and transfer

Human in vitro fertilization (IVF), embryo culture (EC), and embryo transfer (ET) are commonly performed in various media supplemented with blood serum. To determine whether serum is really necessary, the results of IVF, EC, and ET were compared by using two types of media: B2 medium supplemented with human cord serum and B3 medium without any serum. B3 medium is similar to B2, except that it contains 1% pure controlled human serum albumin in place of 1% bovine serum albumin. We did not observe any difference between the results obtained by using B2 or B3 at any phase of IVF, EC, and ET processes. Both media give an overall evolutive pregnancy rate of about 16%. B3 medium with serum seems to increase slightly the cleavage speed. Our results indicate that there are no positive effects when serum is used for human IVF-ET. To avoid serum supplementation is of evident interest for the homogeneity of the results. This will also lead to a better understanding of human early development and control of the egg quality by metabolic analysis of the media following in vitro EC.     

Treatment with piroxicam before embryo transfer increases the pregnancy rate after in vitro fertilization and embryo transfer

OBJECTIVE: To examine the effect of beta-cyclodextrin piroxicam treatment for priming of the uterus on the pregnancy outcome of IVF-embryo transfer (ET) programs. DESIGN: Prospective, randomized, double-blinded placebo-controlled clinical study. SETTING: Large urban medical center. PATIENT(S): One hundred eighty-eight consecutive cycles of fresh IVF-ET and 78 cycles of frozen-thawed ET. The patients underwent IVF because of tubal, male infertility, unexplained, or endometriosis factors. They were randomly divided into treatment and control groups. INTERVENTION(S): In the treatment group, 94 cycles in fresh ET and 39 cycles in frozen-thawed ET the patients received an oral dose of 10 mg of piroxicam. In the control group, the same number cycles corresponding to the treatment group were treated with placebo. Both groups started piroxicam or placebo treatment 1-2 hours before ET. Patients and staff were blinded to the treatment. MAIN OUTCOME MEASURE(S): Implantation rate (IR) and pregnancy rate (PR). RESULT(S): Piroxicam increased significantly IR (18.7%) and PR (46.8%) compared to the control group (8.6% and 27.6%, respectively) in fresh cycles. With the exception of an unexplained factor, patients with the tubal, male infertility, or endometriosis factor had significantly higher PR in the treatment group compared to the control group. The beneficial effect of piroxicam was found in patients less than 40 years old, but was not found in patients more than 40 years. In frozen-thawed cycles, there were statistically significant differences between the treatment group and the control group in IR (9.4% vs. 2.3%) and PR (25.6% vs. 7.7%), respectively. CONCLUSION(S): Our study showed that piroxicam increases IR and PR after IVF-ET in both fresh and frozen-thawed ET cycles. The beneficial effect seems to be more remarkable in patients less than 40 years old with tubal, male infertility, or endometriosis factors. These results suggest that piroxicam treatment before ET is very effective in the priming of a uterus suitable for embryo implantation. This is the first study to investigate the possible consequence of piroxicam for improving the PR after IVF-ET.     

Mouse embryo culture as quality control for human in vitro fertilization: the one-cell versus the two-cell model

Female mice were superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) and mated with male mice. One-cell (n = 429) and 2-cell (n = 450) embryos were collected 20 and 42 hours after hCG and cultured in Ham's F-10 medium (Gibco, Grand Island, NY) (282 mOsm/l, pH 7.4) and in media of altered osmolality (260, 300, 316 mOsm/l), altered pH (7.0, 7.8, 8.0) or various dilutions of Cidex (Surgikos, Arlington, TX) (1:1000, 1:10,000, 1:100,000). Stages of development were observed for 4 days. The development of embryos in the 1-cell system was significantly impaired under all studied conditions by the 4-cell stage of development. The 2-cell system failed to detect trace amounts of Cidex in the culture media and an increase in osmolality to 300 mOsm/l. Other changes in osmolality (260 mOsm/l) and pH (7.8) were detected by the 2-cell system only at the blastocyst stage. The authors conclude that the 1-cell system is more sensitive than the 2-cell system to mild changes in in vitro fertilization culture media.     

In vitro fertilization and embryo transfer

In vitro fertilization is one of the newest approaches to infertility. Opportunities for nurses in IVF involve teaching couples about physiology, providing technical assistance and care, and counseling about the medical and emotional aspects of the program. Many times the nurse serves as the primary support person for couples and as their advocate with other team members.     

体外受精—胚胎移植前切除积水输卵管的意义

目的探讨体外受精-胚胎移植 (in vitro fertilization-embryo transfer,IVF-ET) 前切除积水的输卵 管的意义。方法回顾性分析因输卵管积水导致不孕而要求接受IVF-ET的32例患者,其中17例病人在 IVF-ET切除了积水的输卵管,共接受IVF-ET27个周期,为观察组;有15例病人未切除积水输卵管,共接 受IVF-ET26个周期,为对照组。比较两组病人卵巢对超排卵的反应、受精率、卵裂率、累计胚胎评分(cumula tive embryo score,CES)、胚胎种植率、临床妊娠率。结果观察组受精率、卵裂率、胚胎种植率、临床妊娠率分别 为76.9％、75.0％、11.6％、33.9％,均显著地高于对照组的69.2％、66.7％、2.3％、7.7％,P＜0.05;观察组CES 为(43.4±11.2),非常显著地高于对照组(36.6±13.1),P＜0.01;观察组和对照组的获卵数分别为(13.1±9.7) 个和(12.9±8.2)个,无显著差异。结论IVF-ET前切除积水的输卵管可改善IVF-ET的结局,并且不影响 卵巢对超排卵的反应。     

Cost-effectiveness of in vitro fertilization and embryo transfer

OBJECTIVE: Economic analyses in reproductive medicine often fail to take into account the chances of treatment-independent conception. We compared the cost-effectiveness of several realistic strategies involving IVF using no treatment as the reference strategy. DESIGN: A decision tree was constructed for a subfertile couple in which the clinician had to decide whether to offer treatment with IVF. No treatment at all was used as the reference strategy. SETTING: An analytic decision-making framework. PATIENT(s): Hypothetical subfertile couples. INTERVENTION(s): Two potential treatment approaches: three IVF cycles performed as soon as possible, or no treatment performed initially and then three or four IVF cycles performed if a pregnancy resulting in a live birth does not occur naturally after 212 years. MAIN OUTCOME MEASURE(s): The cost of establishing one pregnancy that results in a live birth. RESULT(s): The cost-effectiveness of IVF depended not only on the costs and expected success rates of IVF itself but also on the couple-specific chances of treatment-independent conception. Consequently, the cost-effectiveness of IVF is strongly dependent on the age of the female partner. This finding corresponds with everyday clinical experience. CONCLUSION(s): Economic analyses in reproductive medicine should take into account the option of providing no treatment.     

Purification of water for in vitro fertilization and embryo transfer

The purpose of this study was to determine whether water obtained from the Milli-Q water purification system (Millipore, Mississauga, Canada) needed further purification for use in in vitro fertilization and embryo transfer. We describe a method for maintenance of the Milli-Q system. To assess water quality, alternate batches of culture media were prepared by using either Milli-Q water or Milli-Q water further treated by twice glass distillation. The percentage of mouse two-cell embryos that developed to blastocysts and the human in vitro fertilization-embryo transfer pregnancy rates were recorded for each batch. There were no significant differences in the parameters examined, indicating that further purification by twice glass distillation is not necessary if the Milli-Q system has been maintained as outlined.     

Influence of embryo transfer depth on in vitro fertilization and embryo transfer outcomes

OBJECTIVE: To investigate the influence of transfer distance from the fundus (TDF) on clinical pregnancy rate (PR) and ectopic pregnancy rate. DESIGN: Retrospective cohort. Between January 2000 and December 2001, 699 ultrasound (US)-guided embryo transfers were conducted. Mock transfer was performed to measure uterine cavity depth 1 month before treatment. Cavity depth was measured by abdominal US before the transfer, from the vaginal stripe to the fundus. Transfers were performed with a Wallace embryo transfer catheter (Cooper Surgical, Shelton, CT) using US and physician's judgment of cavity depth. Transfer distance from the fundus was calculated by subtracting the depth of catheter insertion from the cavity depth, as determined by US or by mock transfer. Statistical analyses were performed by building a multivariable logistic regression model to calculate odds ratios and 95% confidence intervals (CI). SETTING: Women aged 23 to 43 years who are in a university-affiliated, community-based IVF program in Springfield, Massachusetts. PATIENT(S): All patients enrolled in IVF program undergoing embryo transfer. INTERVENTION(S): No patient received any additional procedure or intervention. All of the measurements obtained with the embryo catheter and the transvaginal ultrasound were part of the program's protocol for the embryo transfer. MAIN OUTCOME MEASURE(S): Odds ratio examining relationship between embryo transfer depth and PR.Clinical, implantation, and ectopic PR were 37%, 20%, and 2.1%. Cavity depth by US differed from cavity depth by mock by at least 10 mm in >30% of cases. The TDF by US was highly predictive of PR; TDF by mock was not predictive of PR. Increasing the TDF by US resulted in significantly increased PR as well as lower ectopic rates. Using regression analysis, the odds ratio for TDF by US was 1.11 (95% CI: 1.07-1.14). This suggests that for every additional millimeter embryos are deposited away from the fundus, the odds of clinical pregnancy increased by 11%. CONCLUSION(S): After controlling for potential confounders, the clinical PR is significantly influenced by the transfer distance from the fundus. Cavity depth by US is clinically useful to determine the depth beyond which catheter insertion should not occur.     

胚胎移植位置对体外受精-胚胎移植结局的影响

目的探讨体外受精-胚胎移植（IVF-ET）中,在B超引导下将胚胎移植到子宫腔内的不同位置对妊娠率及胚胎种植率的影响。方法回顾性分析接受IVF-ET治疗的98例输卵管因素不孕患者（共108个周期）的临床资料,根据胚胎移植位置的不同分为两组：A组,胚胎移植到距宫底≥5～〈10mm,共51例患者,56个周期;B组,胚胎移植到距宫底≥10～≤15mm,共47例患者,52个周期。比较两组患者的临床妊娠率、胚胎种植率、异位妊娠率、多胎妊娠率及流产率。结果A组56个周期中,有14个周期妊娠,临床妊娠率为25％,移植胚胎121个,种植17个,胚胎种植率为14％;B组52个周期中,有23个周期妊娠,临床妊娠率为44％,移植胚胎115个,种植28个,胚胎种植率为24％。B组临床妊娠率和胚胎种植率显著高于A组,两组分别比较,差异均有统计学意义（P〈0．05）。两组的多胎妊娠率、异位妊娠率和流产率分别比较,差异均无统计学意义（P〉0．05）。结论在IVF-ET中,胚胎移植位置的不同可影响临床妊娠率和胚胎种植率。     

Monitoring gonadotropin-stimulated cycles for in vitro fertilization and embryo transfer

One hundred fifty-one cycles stimulated with human menopausal gonadotropin (hMG) and/or folliclestimulating hormone (FSH) were analyzed retrospectively to determine the relationship of the monitoring parameters used: serum estradiol (E₂), transvaginal sonographic follicular size, and peripheral biologic estrogen response, with harvests and pregnancies resulting from in vitro fertilization and embryo transfer. Mean ± SE serum E₂ levels were higher on the day of human chorionic gonadotropin (hCG) administration in the miscarriage group (689.4±27.5 pg/ml) than in the nonpregnant group (527.7±25.8 pg/ml) (P=0.018) and highest in the ongoing pregnancy group (734.6±66.5 pg/ml) (P=0.003). When two or more preovulatory oocytes (preovs) were retrieved, the mean E₂ levels were higher (622.3±27.3 pg/ml) than if zero or one preov was retrieved (378.4±43.5 pg/ml) (P=0.001). The mean diameter of the two largest follicles at the time of hCG administration was significantly greater (14.7 mm) in the group with two or more preovs retrieved than in the group with zero or one preov (13.3 mm) (P=0.001). The relative probability of achieving a pregnancy was best predicted by the presence of two follicles16 mm in diameter on transvaginal ultrasound examination and was 1.63 times greater than that of all patients in this series.     

Hysteroscopic cervical canal shaving: a new therapy for cervical stenosis before embryo transfer in patients undergoing in vitro fertilization

OBJECTIVE: To report a case of cervical stenosis repaired by hysteroscopic cervical shaving that created a smooth passage for ET in a patient undergoing IVF. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A woman with a history of multiple failed IVF attempts in whom ET was extremely difficult. INTERVENTION(S): Operative hysteroscopy with creation of a cervical tract. MAIN OUTCOME MEASURE(S): Ease of postoperative ET and outcome of IVF treatment after the hysteroscopic procedure. RESULT(S): Easy performance of ET and a resulting triplet pregnancy. CONCLUSION(S): This novel hysteroscopic repair of cervical stenosis resulted in a markedly easier ET and a viable pregnancy.     

输卵管栓塞术在体外受精-胚胎移植前处理输卵管积水中的应用

目的 探讨输卵管栓塞术在体外受精-胚胎移植(ⅣF-ET)前处理输卵管积水中的应用.方法 选择因输卵管积水行输卵管栓塞术的不孕症患者46例(栓塞组)和同期双侧输卵管阻塞的不孕症患者91例(对照组),栓塞组行输卵管栓塞术,将微弹簧圈通过微导管送入输卵管间质部和峡部,1个月后行子宫输卵管造影术检查栓塞效果.栓塞术后3个月,两组患者行IVF-ET治疗,观察并比较每个周期内的平均获卵数、受精率、临床妊娠率、异位妊娠率、流产率等各项指标.结果 栓塞组共栓塞82条输卵管,插管均1次成功.栓塞效果显著的输卵管有72条,占88%;有效10条,占12%;无效为0.栓塞组患者的平均获卵数(18.9个)、受精率(69%)、临床妊娠率(41%)均高于对照组(分别为17.6个、63%、39%),但差异无统计学意义(P>0.05);输卵管妊娠率(0)、流产率(8%)均低于对照组(分别为8%、16%),差异有统计学意义(P<0.05).结论 输卵管栓塞术应用于IVF-ET前对输卵管积水进行处理,是一种简便、安全、经济的方法,有增加临床妊娠率的趋势,并且能杜绝输卵管妊娠的发生,是一种可行且有效的方法.     

Mock embryo transfer in early luteal phase, the cycle before in vitro fertilization and embryo transfer: a descriptive study.

OBJECTIVE: To investigate visually the uterine retention or sequestration of boluses of radiopaque dye, mimicking embryo transfer (ET). DESIGN: During the cycle before in vitro fertilization (IVF) and ET, patients underwent a mock ET of 40 microL of radiopaque dye into the uterine cavity. The patient was positioned supine for retroverted or axial and knee chest for anteverted uteri. The position of the dye at injection, during and after catheter removal, and during patient roll over and standing was monitored. SETTING: Treatment of infertility in a private practice. PATIENTS: Thirty-four IVF patients. INTERVENTIONS: During the cycle before IVF/ET, patients underwent a mock ET using a bolus of 40 microL of radiopaque dye. MAIN OUTCOME MEASURES: Planned before visual observation began. RESULTS: The dye remained primarily in the uterine cavity in only 68% (optimum ET position is knee chest for anteverted and supine for retroverted or axial uterus) and 48% (nonoptimum position is supine for anteverted uterus) at mock ET; in those groups, a 33% clinical pregnancy rate (PR) per retrieval resulted. Dye motility into the fallopian tube(s), cervix, and/or vagina is 38.2%, 8.8%, and 11.8%, respectively. CONCLUSIONS: If the mock ET had been the actual ET, 32% (optimum ET position) and 52% (nonoptimum ET position) of all patients would have lost their opportunity for pregnancy as a result of the ET procedure. Our 33% PR per retrieval among those patients who retained the dye in utero is more consistent with our expectations, given the advanced technologies of IVF/ET today.