A novel technique for oviduct occlusion to generate live births from cryopreserved rabbit oocytes after in vivo fertilisation.

Intraoviductal transfer technique in combination with in vivo fertilisation has arisen as an effective technique to assess live births after transfer of slow-frozen oocytes in the rabbit. Nevertheless, the great disadvantage of this method is the accumulation of tubal fluid in a large number of females after clamping the oviducts. In this study, we develop an alternative method to minimise damage to the oviduct and increase the birth rate. The aims of this study were (1) to evaluate the ability of cyanoacrylate tissue adhesive to occlude the oviduct for female sterilisation; (2) to evaluate the effect of oviduct occlusion immediately after transferring fresh oocytes on in vivo fertilisation; and (3) to assess this technique to generate live births from fresh and slow-frozen oocytes. In all the experiments, recipients were artificially inseminated 9h prior to occluding the oviducts. In the first experiment, the left oviduct was blocked with cyanoacrylate tissue adhesive, while the right one was used as a control. Six days later, oviducts and uterine horns were flushed to assess embryo recovery rates. While the embryo recovery rate was 79.2% in the intact oviduct, no embryos were recovered in the blocked one. In the second experiment, fresh oocytes were transferred into both oviducts, which were immediately occluded. Six days later, the in vivo fertilisation success rate was 33.7%. Finally, in the last experiment, slow-frozen oocytes were transferred and the rate of live births was 13.2±4.5%. The study shows that when using this method the generation of live births from slow-frozen oocytes increases significantly. In addition, our results suggest that in vivo environment could help improve the results of oocyte cryopreservation.

Intrauterine insemination versus fallopian tube sperm perfusion for non-tubal infertility.

BACKGROUND: Intrauterine insemination (IUI) is a common treatment for couples with subfertility that does not involve the fallopian tubes. It is used to bring the sperm close to the released oocyte. Another method of introducing sperm is fallopian tube sperm perfusion (FSP). Fallopian tube sperm perfusion ensures the presence of higher sperm densities in the fallopian tubes at the time of ovulation than does standard IUI. These treatments are often used in combination with ovarian hyperstimulation. OBJECTIVES: To compare intrauterine insemination versus fallopian tube sperm perfusion in the treatment of non-tubal subfertility, for live birth and pregnancy outcomes. SEARCH METHODS: We searched the Menstrual Disorders and Subfertility Group Trials Register, MEDLINE, CINAHL and EMBASE from inception to September 2013. We also searched study reference lists and trial registers. SELECTION CRITERIA: Randomised controlled trials (RCTs) comparing IUI with FSP in couples with non-tubal subfertility were included. DATA COLLECTION AND ANALYSIS: Two review authors independently selected studies for inclusion, assessed study quality and extracted the data. If studies were sufficiently similar, data were combined using a fixed-effect model to calculate pooled odds ratios (ORs) and 95% confidence intervals (CIs). A random-effects model was used if substantial statistical heterogeneity was detected. Studies that included participants with unexplained or mixed (non-tubal) subfertility were analysed separately from studies restricted to participants with mild or moderate male factor subfertility. The overall quality of evidence for the main outcomes was summarised using Grading of Recommendations Assessment, Development and Evaluation (GRADE) criteria. MAIN RESULTS: The review included 16 RCTs. Fourteen RCTs (1745 women) were included in the meta-analysis. Only three studies reported live birth per couple. No evidence of a statistically significant difference was noted between IUI and FSP in live birth (OR 0.94, 95% CI 0.59 to 1.49, three RCTs, 633 women, I(2) = 0%, low-quality evidence) or clinical pregnancy (OR 0.75, 95% CI 0.49 to 1.12, 14 RCTs, 1745 women, I(2) = 52%, low-quality evidence). These findings suggest that for a couple with a 13% chance of live birth using FSP, the chance when using IUI will be between 8% and 19%; and that for a couple with a 19% chance of pregnancy using FSP, the chance of pregnancy when using IUI will be between 10% and 20%. Nor was evidence found of a statistically significant difference between IUI and FSP in per-pregnancy of multiple pregnancy (OR 0.96, 95% CI 0.44 to 2.07, eight RCTs, 197 women, I(2) = 0%, low-quality evidence), miscarriage (OR 1.23, 95% CI 0.60 to 2.53, seven RCTs, 199 women, I(2) = 0%, low-quality evidence) or ectopic pregnancy (OR 1.71, 95% CI 0.42 to 6.88, four RCTs, 111 women, I(2) = 0%, very low quality evidence). Substantial heterogeneity was noted for the outcome of clinical pregnancy (I(2) = 54%), for which no clear explanation was provided. AUTHORS' CONCLUSIONS: Currently no clear evidence suggests any difference between IUI and FSP with respect to their effectiveness and safety for treating couples with non-tubal subfertility. However, a high level of uncertainty is evident in the findings, and additional research may be useful.

Successful pregnancy outcome following gamete intra-Fallopian transfer in a patient with Müllerian dysgenesis.

A 29-year-old lady with Müllerian dysgenesis was keen to have a baby. Clinically, she was medium built with well-developed secondary female sexual characteristics. There was a short and blind vagina. She had undergone surgery for an imperforated hymen. Her FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent gamete intra-Fallopian transfer (GIFT). Oocyte retrieval was carried out laparoscopically and a total of nine oocytes were retrieved. Four of the oocytes were transferred together with motile spermatozoa into the right Fallopian tube and the remaining five oocytes were inseminated with spermatozoa for IVF. Three embryos resulted and were frozen. She subsequently developed moderate ovarian hyperstimulation syndrome. Serum ß-human chorionic gonadotrophin concentration 14 days after GIFT was 1612 IU/l. Her antenatal care was relatively uneventful until 31 weeks of gestation when she was diagnosed to have intrauterine growth retardation and oligohydramnios. She then underwent an emergency Caesarean section at 32 weeks of pregnancy delivering a normal baby. This case study describes a successful pregnancy outcome following gamete intra-Fallopian transfer (GIFT) in a woman with malformation of the vagina (Müllerian dysgenesis). A 29-year-old lady with Müllerian dysgenesis diagnosed at 16 years of age was keen to become pregnant. Upon examination, a decision was made for a William's vulvovaginoplasty but as the patient was indecisive the surgery was deferred. Clinically, she is a medium-built lady with well-developed secondary female sexual characteristics. There was a short and blind vagina. Her serum FSH and LH concentrations were normal. Laparoscopy revealed a patent right Fallopian tube, a rudimentary right uterus and extensive pelvic endometriosis. She subsequently underwent GIFT. Nine oocytes were retrieved through laparoscopy. Four of the oocytes were transferred together with motile sperm into the right Fallopian tube and the remaining five oocytes were inseminated with sperm for IVF. Three embryos resulted and were frozen. Serum ß human chorionic gonadotrophin concentration measured 14 days after GIFT was 1612 IU/l. An abdominal ultrasonography performed at 5 weeks showed one intrauterine gestational sac. Her antenatal care was uneventful until 31 weeks of gestation when she developed a deficiency of amniotic fluid in the amniotic sac. She then underwent an emergency Caesarean section at 32 weeks of pregnancy. She delivered a healthy, normal 1.24 kg baby boy. Her post-natal care was uneventful.

Gamete Intrafallopian Transfer Versus Super-Ovulation with Intrauterine Insemination for the Treatment of Infertility : a Prospective Randomised Study on Pregnancy Outcome

Background. A prospective randomised controlled trial comparing gamete intrafallopian transfer (GIFT) with intrauterine insemination (IUI) was undertaken at the Fertility and Reproductive Biology Unit of the Department of Obstetrics and Gynaecology; Tygerberg Hospital; between July 1999 and June 2000. Method. Eighty-five women were included the study and were randomly allocated between the two groups after routine infertility investigations; 41 women to IUI and 44 women to GIFT. A combination of clomiphene incitrate and human menopausal gonadotropin was administered to both groups to achieve ovulation. Results. Six (13.6) of the 44 cycles in the IUI group and 24 (53.3) of the 45 cycles in the GIFT group achieved conception. The mean number of cycles needed to achieve pregnancy in IUI was 7.3 (44/6) and in GIFT was 2.05 (45/24). The ongoing pregnancy rate of GIFT was 39.7 more effective than that of IUI (p=0.0001.The total ongoing pregnancy rate of GIFT was 30.8 superior to that of IUI (p=0.0021). When 2 folli-cles were obtained in an IUI cycle; GIFT was 41.6 more effective (p=0.0024); and when more than 2 follicles were obtained; GIFT was 28.3 more effective (p=0.0265). Conclu- sions. The number of mature follicles significantly increased the chance of pregnancy with IUI. In comparing the number of cycles needed to achieve a pregnancy; 1 GIFT cycle is equivalent to more than 3 IUI cycles. It is important to note that 4 IUI cycles will give equivalent or even better results if 2 - 3 follicles are recruited per cycle. In spite of the greater efficacy of GIFT; the authors conclude that at least 3 to 4 IUI cycles should be attempted before GIFT; on the basis that it is more cost effective and less invasive

Comparison of unilateral and bilateral transfer of injected oocytes into fallopian tubes: a prospective, randomized clinical trial.

OBJECTIVE: To assess whether pregnancy rate differs in unilateral vs. bilateral transfer in "immediate transfer of injected oocytes into the fallopian tubes." DESIGN: Prospective randomized clinical trial. SETTING: Tertiary university hospital. PATIENT(S): The study population included 160 patients presenting with male factor infertility who fulfilled the criteria for immediate transfer of injected oocytes into tubes. INTERVENTION(S): Four injected oocytes were transferred into two tubes (study group) or one tube (control group). MAIN OUTCOME MEASURE(S): Implantation rate, clinical pregnancy rate, multiple pregnancy rate, and incidence of pregnancy with unknown location (PUL). RESULT(S): A total of 72 (45%) pregnancies were achieved. There were 32 pregnancies (1 PUL, 4 multiple, and 27 singletons) in the bilateral transfer group and 40 pregnancies in the unilateral transfer group (1 PUL, 7 multiple, and 32 singletons). No significant difference was found in the implantation rate, clinical pregnancy rate, multiple pregnancy rate, and incidence of PUL. CONCLUSION(S): This study demonstrates that no difference in outcome occurred between unilateral and bilateral transfer in microinjected oocytes intrafallopian transfer (MIFT). Therefore, along with the same outcome parameters, unilateral transfer is the preferred method of MIFT.

Xenogenous fertilization of equine oocytes following recovery from slaughterhouse ovaries and in vitro maturation.

The in vitro production (IVP) of equine embryos using currently available protocols has met limited success; therefore investigations into alternative approaches to IVP are justified. The objective of this study was to evaluate the feasibility of xenogenous fertilization and early embryo development of in vitro matured (IVM) equine oocytes. Follicular aspirations followed by slicing of ovarian tissue were performed on 202 equine ovaries obtained from an abattoir. A total of 667 oocytes (3.3 per ovary) were recovered from 1023 follicles (recovery rate, 65%). Oocytes underwent IVM for 41 +/- 2 h (mean +/- S.D.), before being subjected to xenogenous gamete intrafallopian transfer (XGIFT). An average of 13 +/- 0.8 oocytes and 40x10(3) spermatozoa per oocyte were transferred into 20 oviducts of ewes. Fourteen percent of transferred oocytes (36/259) were recovered between 2 and 7 days post-XGIFT and 36% of those recovered displayed embryonic development ranging from the 2-cell to the blastocyst stage. Fertilization following XGIFT was also demonstrated by the detection of zinc finger protein Y (ZFY) loci. Ligation of the uterotubal junction (UTJ), ovarian structures, or the duration of oviductal incubation did not significantly affect the frequency of embryonic development or recovery of oocytes/embryos after XGIFT. In conclusion, equine embryos can be produced in a smaller non-equine species that is easier for handling.

Gamete intra-fallopian transfer or intrauterine insemination after controlled ovarian hyperstimulation for treatment of infertility due to endometriosis.

We compared the effectiveness of gamete intra-Fallopian transfer (GIFT) and intrauterine insemination (IUI) after controlled ovarian hyperstimulation (COH) in the treatment of infertility due to endometriosis. This was a retrospective study carried out at a tertiary teaching medical center. A total of 127 consecutive patients with endometriosis were treated with GIFT or IUI after COH between June 1990 and December 1998. Patients were divided into two groups. Group 1 (n = 97) included patients with stages 1 and 2 endometriosis, and group 2 (n = 30) included patients with stages 3 and 4 endometriosis. Laparoscopic conservative surgery for endometriosis was performed prior to IUI for patients in both group 1 and group 2. In group 1, 55 patients underwent 95 cycles of IUI after COH and 42 patients underwent 57 cycles of GIFT. In group 2, 14 patients underwent 16 cycles of IUI after COH, while 16 patients underwent 22 cycles of GIFT. The stimulation protocol for both GIFT and IUI was mid-luteal pituitary down-regulation with a gonadotropin releasing hormone agonist (GnRH-a) followed by gonadotropins. In group 1, the pregnancy rates (GIFT = 50.9%, IUI = 29.4%) and the delivery rates (GIFT = 28.1%, IUI = 14.7%) per cycle were significantly higher in GIFT compared to IUI (p = 0.009 and p = 0.05, respectively). There was no significant differences in the pregnancy rate (GIFT 69%, IUI 50.9%, respectively) or the delivery rate (GIFT 38.1%, IUI 25.5%) per patient. In group 2, there was no significant difference in the pregnancy rate (GIFT 54.5%, IUI 31.3%) or the delivery rate (GIFT 40.9%, IUI 12.5%) per cycle, but the difference in the pregnancy rate (GIFT 75%, IUI 35.7%) and the delivery rate (GIFT 56.3%, IUI 14.3%) per patient was significantly higher in GIFT compared to IUI (p = 0.04 and p = 0.02, respectively). We conclude that, when the same stimulation protocol is used in the early stages of endometriosis, a few cycles of IUI can achieve similar results to GIFT, and therefore should be used first. In advanced stages of endometriosis GIFT appears to be more effective.

Equine sperm-oocyte interaction: results after intraoviductal and intrauterine inseminations of recipients for oocyte transfer.

Insemination of recipients for oocyte transfer and gamete intrafallopian transfer (GIFT) in five experiments were reviewed, and factors that affected pregnancy rates were ascertained. Oocytes were transferred into recipients that were (1) cyclic and ovulated at the approximate time of oocyte transfer, (2) cyclic with aspiration of the preovulatory follicle, and (3) noncyclic and treated with hormones. Recipients were inseminated before, after, or before and after transfer. Intrauterine and intraoviductal inseminations were done. Pregnancy rates were not different between cyclic and noncyclic recipients (8/15, 53% and 37/93, 39%). The highest numerical pregnancy rates resulted when recipients were inseminated with fresh semen from fertile stallions before oocyte transfer or inseminated with cooled transported semen before and after oocyte transfer. Oxytocin was administered to recipients before oocyte transfer when fluid was imaged within the uterus. Administration of oxytocin to recipients at the time of oocyte transfer resulted in significantly higher pregnancy rates than when oxytocin was not administered (17/26, 65% and 28/86, 33%). Intraoviductal and intrauterine inseminations of recipients during oocyte transfer resulted in similar embryo development rates when fresh semen was used (12/22, 55% and 14/26, 55%). However, embryo development rates significantly reduced when frozen (1/21, 5%) versus fresh sperm were inseminated into the oviduct. Results suggest that insemination of a recipient before and after transfer could be beneficial when semen quality is not optimal; however, a single insemination before transfer was adequate when fresh semen from fertile stallions was used. Absence of a preovulatory follicle did not appear to affect pregnancy rates in the present experiments. The transfer of sperm and oocytes (GIFT) into the oviduct was successful and repeatable as an assisted reproductive technique in the equine.

Conscious sedation versus general anaesthesia for minilaparoscopic gamete intra-Fallopian transfer: a prospective randomized study.

BACKGROUND: Gamete intra-Fallopian transfer (GIFT) is a successful technique for infertile women, and is performed almost exclusively by laparoscopy under general anaesthesia. METHODS: We performed a prospective randomized study of 67 infertile patients to assess the efficacy of minilaparoscopic GIFT under conscious sedation/local anaesthesia (group A) compared with general anaesthesia (group B). Operative and discharge times and pregnancy outcome were evaluated in both groups. RESULTS: Operative time was similar in both groups. The rate of patients discharged 2 h after surgery was significantly higher in group A. The necessity for postoperative analgesics was significantly higher in group B. No significant differences were noted between the two study groups in terms of pregnancy outcome. CONCLUSIONS: Conscious sedation and local anaesthesia will allow us to perform an outpatient minilaparoscopic GIFT without the need for general anaesthesia. The simplicity of the method, and the quicker discharge time in comparison with general anaesthesia, offer a detectable benefit for patients.

Comparison of culture and insemination techniques for equine oocyte transfer.

This study was designed to test 3 approaches for insemination and transfer of oocytes to recipient mares. Oocytes were recovered transvaginally from naturally cycling donor mares 24 to 26 h after an intravenous injection of 2500 IU of hCG when follicles reached 35 mm in diameter. Multiple oocytes (1 to 4) were transferred surgically into the oviducts of 4 or 5 recipient mares per group. Three groups of transfers were compared: 1) transfer of oocytes cultured in vitro for 12 to 14 h postcollection with insemination of the recipient 2 h postsurgery; 2) transfer of oocytes into the oviduct within 1 h of collection, with completion of oocyte maturation occurring within the oviduct, and insemination of the recipient 14 to 16 h postsurgery; and 3) transfer of spermatozoa and oocytes (cultured 12 to 14 h in vitro) into the oviduct. Numbers of embryos detected by Day 16 of gestation were not different (P>0. 1) for groups 1, 2, and 3 (57%, 43% and 27%). Therefore, equine oocytes successfully completed the final stages of maturation within the oviduct, and sperm deposited within the oviduct were capable of fertilizing oocytes.

Treatments resulting in pregnancy in nonovulating, hormone-treated oocyte recipient mares.

Synchronization of follicle growth between oocyte donor and recipient mares is difficult. To avoid this, recipient mares in a clinical program were used during a period of low follicular activity, and were treated with estrogen before transfer and progesterone after transfer. Five pregnancies were established after oocyte transfer to nonovulating, hormone-treated recipient mares. One pregnancy was lost before 30 d gestation, and the other 4 foals were carried to term. One foal died at birth. Establishment and maintenance of pregnancy in these mares indicates that nonovulating, hormone-treated mares may offer an alternative to cyclic recipients in oocyte transfer programs.

Effect of timing of follicle aspiration on pregnancy rate after oocyte transfer in mares.

Mares with preovulatory follicles >33 mm in diameter were administered hCG and were randomly assigned for aspiration of the dominant follicle at 24 h or 35 h after hCG administration. Oocytes recovered at 24 h were cultured for 12 h before transfer and oocytes recovered at 35 h were cultured for 1 h. Oocytes were transferred by flank laparotomy to the oviduct of the same mare, or to the oviduct of another oocyte donor. Recipient mares were inseminated before and after transfer. The oocyte recovery rates at 24 h and 35 h after hCG administration were not significantly different (10/15 (66%) and 11/15 (73%), respectively) and resulted in an overall recovery rate of 70%. The overall pregnancy rate after transfer was 9/17 (53%) and there was no significant difference between groups (5/8 (63%) for the 24 h group and 4/9 (44%) for the 35 h group). The presence of uterine fluid in recipient mares > 2 days after transfer was associated with a significantly lower pregnancy rate (3/10 versus 6/7 for mares that had no fluid after day 2). This study indicates that the timing of oocyte collection after administration of hCG is not a major determinant of the pregnancy rate after oocyte transfer. Medication associated with follicle aspiration and oocyte transfer may increase susceptibility of recipient mares to endometritis, which can lower pregnancy rates if not resolved.

Oviductal insemination of mares.

A technique was developed for oviductal insemination of mares, in which a small number of motile spermatozoa are deposited directly into the oviduct. Pregnancy rates in mares inseminated by traditional intrauterine artificial insemination were compared with rates in mares inseminated by oviductal insemination. Fifteen mares were inseminated with 5 x 10(8) progressively motile spermatozoa by intrauterine artificial insemination, and 14 mares were inseminated with 5 x 10(4) progressively motile spermatozoa by oviductal insemination. Pregnancy rates in mares inseminated by intrauterine artificial insemination (40%) and oviductal insemination (21.4%) were not significantly different (P > 0.05). This study indicates that oviductal insemination can produce pregnancies in mares using 10,000 times fewer spermatozoa than are used for intrauterine artificial insemination.

[Tubal catheterization for transvaginal gamete intrafallopian transfer: an alternative technique for non-tubal infertility].

OBJECTIVE: To evaluate the outcome of transvaginal gamete intrafallopian transfer (TV-GIFT) for infertile patients with non-tubal factors. METHODS: Controlled ovarian hyperstimulation was used for all patients. Transvaginal ultrasound-guided oocyte retrieval followed by tubal catheterization for transvaginal gamete intrafallopian transfer of 3.3 +/- 2.1 oocytes (range 1-8) and 200,000 sperms in 50 microliters culture medium. RESULTS: The TV-GIFT was used in total of 23 treatment cycles. Of 23 cycles performed for TV-GIFT, the procedure was completed easily in 18 and difficult in 5, resulting in 5 clinical pregnancies(21.7% per cycle), 2 of which resulted in abortion, 2 ongoing pregnancy one was ectopic pregnancy. No pregnancy occurred in four cycles with only one oocyte transferred. CONCLUSIONS: Simple and cost-effective TV-GIFT may achieve a higher pregnancy rate. Compared with laparoscopically directed GIFT, tubal catheterization for TV-GIFT is safe and less invasive. It is an alternative for non-tubal infertility before entering the IVF-ET program.

[Transvaginal gamete intrafallopian transfer: an analysis of 42 cases].

OBJECTIVE: To develop the technique of transvaginal gamete intrafallopian transfer (TV-GIFT) and to evaluate its effectiveness. METHODS: From May, 1996 to October, 1997, 42 infertile patients with at least one tube patent, proven by hysterosalpingography were included. In 43 cycles, ovarian stimulation was performed by using follicle-stimulating hormone (FSH), human menopausal gonadotropin (hMG)/hCG or hMG/hCG program. Oocytes retrieval was performed by transvaginal ultrasound guided needle aspiration. Semen samples were prepared by modified swim-up method. The Jansen-Andersen tubal catheter was introduced into a fallopian tube transvaginally, and the gametes were injected into the tube. RESULTS: Transvaginal cannulation was successfully performed in 67.4% of cases. The mean number of oocytes transferred was 4.0 +/- 0.9 (2-6) per cycle. 13 cases obtained clinical pregnancy (30.2% per cycle), 9 given birth with 5 twin pregnancies (altogether 13 babies), 4 spontaneous abortions. CONCLUSIONS: TV-GIFT offers an acceptable pregnancy rate for patient with at least one patent tube. The procedure is simple and easy to perform and therefore, is worth to recommend for the treatment of infertility.

Gamete intrafallopian transfer. Comparison of epidural vs. general anesthesia.

OBJECTIVE: To investigate the reproductive outcome of laparoscopic gamete intrafallopian transfer (GIFT) performed under epidural vs. general anesthesia. STUDY DESIGN: Retrospective analysis of 110 consecutive laparoscopic GIFT procedures performed under general or epidural anesthesia. All patients underwent controlled ovarian hyperstimulation employing human menopausal gonadotropin and gonadotropin releasing hormone agonist, given either in the midluteal or early follicular phase. Data were retrieved concerning age, diagnosis, estradiol levels and maximum follicular diameter at the time of human chorionic gonadotropin injection, percentage of mature oocytes retrieved and number of oocytes transferred. These variables were compared using the independent means t test. Pregnancy rates and outcome between the general and epidural anesthesia groups were compared with the chi 2 test. RESULTS: Of the 110 procedures, 84 were performed under general anesthesia, whereas 22 were done using epidural anesthesia. Four procedures started with epidural anesthesia and were converted to general anesthesia because of upper abdominal discomfort. The success rate of epidural anesthesia, therefore, was 85% (22/26). There were no significant differences in the confounding variables between the general and epidural anesthesia groups. However, patients receiving epidural anesthesia had a significantly higher pregnancy rate, 59.1%, and a live birth rate of 40.9% as compared to 31.0% and 21.4%, respectively, for the general anesthesia group. CONCLUSION: Laparoscopic GIFT can be performed safely under epidural anesthesia. Because of the higher pregnancy and live birth rates, epidural is the anesthetic of choice for GIFT. However, a prospective, randomized study is needed to confirm the above observation.