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[Assisted hatching--a useful micromanipulation technic in women after repeated failure of embryo transfer].

One reason for the low implantation rate after IVF/ET is the limiting ability of blastocysts to escape from the zona pellucida, and of nidation. This procedure can be assisted by the creation of an artificial opening in the zona pellucida before the transfer (assisted hatching). An artificial slot in the zona of 58 embryos (20 patients) was created mechanically by using Leitz Fluovert microscope and Narishige micromanipulators. From 20 embryo transfers 6 clinical pregnancies resulted (30.0% clin. preg. rate). In 4 cases women with three or more unsuccessful transfers were involved. This method should be used in classical IVF, specially in cases of repeated high quality embryo transfers which failed.

Embryo Transfer↗

Risk evaluation for bovine embryo transfer services using computer simulation and economic decision theory.

A mathematical model was used to evaluate the dynamics and risks in bovine embryo transfer. Variables included embryo collection, fertilization, and transfer rates, plus overall pregnancy rates. Decision analysis was applied to three sets of 500 simulated flushes to test three strategies: 1) nonsuperovulation, 2) superovulation using follicle stimulating hormone (FSH) and 3) superovulation using pregnant mare serum gonadotropin (PMSG). Model validation involved comparing model performance against standards derived from 39 published studies. The model's repeatability was +/- 0.10 against standards in 93% of cases, and never exceeded +/- 0.13 in all 1500 simulations. Model outcomes were accurate to +/- 3% using average results. No pregnancies occurred in 22% of nonsuperovulated donors, 8% of FSH superovulated donors, and 6% of PMSG superovulated donors. PMSG treatment averaged more pregnancies per flush than FSH treatment (4.4 vs 3.9) but showed greater variation in response (64 vs 51%). Decision analysis suggests that a PMSG-induced flush would net $105 more than an FSH-induced flush, and that either superovulation strategy would yield approximately 10 times the net income of a nonsuperovulated flush. Pricing by response (PMSG) or by transfer (FSH) is optimal for the provider of embryo transfer services.

Journal Article↗

Ovarian response, embryo recovery and results of embryo transfer in a Hungarian native pig breed.

The objective of the study was to use embryo transfer (ET) for propagation of the Swallow Belly Mangalica population. Mangalica is a native Hungarian pig breed adapted to extreme climate and housing conditions and distinguished for excellent meat and fat quality. However, due to their weak reproductive characteristics and relatively high fat proportion, Mangalica pigs have been replaced by modern breeds. Now, there is an increased interest again to safeguard the properties of this breed. We conducted two experiments. First, we used a total of 18 puberal Mangalica gilts to determine an optimal superovulatory treatment. Following estrus synchronization with Regumate, we injected gilts with either 750, 1000 or 1250 IU PMSG, followed by 750 IU hCG 80 h later. We scanned ovaries endoscopically 3 days after hCG administration. The application of 1000 and 1250 IU PMSG resulted in a higher rate of ovulation compared to 750 IU (24.2 +/- 3.6 and 21.0 +/- 2.3 vs. 13.7 +/- 2.7 P<0.05). The number of follicular cysts increased after administration of 1250 IU PMSG compared to 750 and 1000 IU (2.0 +/- 1.3 vs. 0.3 +/- 0.7 and 0.2 +/- 0.3, P<0.05). Thus, we chose 1000 IU PMSG for further stimulation of Mangalica gilts. In the second experiment, we induced superovulation in 10 Mangalica donor gilts by 1000 IU PMSG and 750 IU hCG. Gilts were fixed-time inseminated, and then five days later embryo collection was carried out surgically (n=6) or endoscopically (n=4). Out of the 187 ova recovered, 92.5% were at the morula/blastocyst stage. The embryo recovery rate was higher following surgical flushing than following endoscopy (91.5 +/- 4.4% vs. 71.4 +/- 12.7%, P<0.05). Altogether 143 embryos were transferred surgically or endoscopically into 8 Landrace recipients. Surgical and endoscopic transfer of Mangalica embryos into Landrace gilts resulted in pregnancies in 3 and 2 gilts, respectively; thus the overall farrowing rate was 62.5%. The birth of 59 Mangalica piglets from 5 embryo recipients equals an average litter size of 11.8 +/- 1.3, which is two times larger than usual in this breed. Therefore, we concluded that an appropriate inter-breed ET program is a suitable tool to propagate the endangered Mangalica breed.

Animals↗

Influence of position and length of uterus on implantation and clinical pregnancy rates in IVF and embryo transfer treatment cycles.

In a prospective study of 807 consecutive women shown to have an apparently normal uterus after hysterosalpingography, hysteroscopy or pelvic ultrasonography prior to IVF or intracytoplasmic sperm injection (ICSI) and embryo transfer, the position and length of the uterine cavity was measured routinely at a pre-treatment mock transfer procedure. The apparent length of the uterine cavity was <7 cm in 128 women (group 1), 7-9 cm in 594 women (group 2) and >9 cm in 85 women (group 3). The uterus was noted to be retroverted in 38. 2% (308) women. The embryo transfer catheter was advanced to 5 mm from the uterine fundus based on the previously determined cavity length in all the embryo transfer procedures at 48 h after oocyte collection. Implantation and clinical pregnancy rates were not significantly different with respect to position of the uterus, difficulties encountered in passage of the catheter, mean age of the women, aetiology or duration of infertility or embryology events. An apparently greater cavity length was seen in older and/or parous women, but the difference was not statistically significant. Although the highest implantation and clinical pregnancy rates were seen in women with a cavity length of 7-9 cm (group 2) the differences were not statistically significant: group 1, 18.9 and 36. 7%; group 2, 21.0 and 46.5%; and group 3, 17.3 and 32.9% respectively. The incidence of ectopic pregnancy per reported clinical pregnancy was highest in group 1 women, being 14.9% (7/47) in comparison with group 2 (1.8%, 5/276) and group 3 (0%, 0/27) (P: < 0.0005), suggesting that the size of the uterus is a critical factor in the aetiology of ectopic pregnancy in IVF/ICSI-embryo transfer.

Adult↗

Induction of twinning in crossbred heifers by ipsilateral frozen embryo transfer.

Induction of twinning by means of nonsurgical ipsilateral transfer of two frozen embryos or of demi-embryos was attempted in 297 virgin crossbred dairy heifers that were kept at the following locations: in stables at our breeding station (Group 1), in a mountain field (Group 2) and at a private farm (Group 3). Of these, 187 heifers (63%) were diagnosed pregnant at 25 d after embryo transfer and 134 heifers (45%) were diagnosed pregnant at 60 d after transfer by ultrasonic echography or by palpation per rectum. At calving, 118 heifers produced 46 sets of twins and 72 single calves, for a total of 164 calves sixteen pregnant heifers aborted fetuses between 2 and 7 mo. The pregnancy rate of Group 1 (39%) was lower than that of Group 2 (48%) or Group 3 (50%). Abortion and mortality rates for Group 3 (8% and 8%, respectively) were significantly lower (P<0.05) than for Group 1 (16% and 12%) or Group 2 (13% and 18%). Twin calves had lower birth weights (P<0.05) than singles, but at 270 to 330 d of age, there was no significant difference between the weights of single and twin calves. Dystocia or difficult delivery was not observed heifers producing twins. Retention of the placenta was observed in twin calving heifers, with placenta retention levels the highest in Group 2 (33%) than followed by Group 1 (14%) and Group 3 (10%). When gestation periods were compared between single and twin calves in Groups 1 and 2, the gestation period of single calves was 5.0 to 7.0 d shorter than that of twin calves. No significant difference was observed between the gestation periods of single and twin births in Group 3.

Journal Article↗

DNA contamination of reagents used in embryo transfer and culture.

Commonly used reagents in the culture and transfer of embryos are isolated from blood and tissue samples and thus have the potential for chromosomal and or mitochondrial DNA contamination. In this study, we evaluated the results obtained from PCR analysis of bovine trypsin, bovine sera, and bovine albumin precipitates. Bovine sera samples that were tested yielded minor to heavy DNA contamination signals depending on the manufacturer and specific type of sera. Bovine albumin precipitates showed very little DNA contamination or none at all. Bovine trypsin samples yielded moderate DNA contamination signals depending on the ability of the trypsin to be inactivated prior to PCR analysis.

Journal Article↗

Is the outcome of in-vitro fertilization and embryo transfer treatment improved by spontaneous or surgical drainage of a hydrosalpinx?

A pilot study was designed to examine whether the outcome of embryo transfer in women with a hydrosalpinx might be improved by surgical drainage of the hydrosalpinx at the time of oocyte collection for in-vitro fertilization treatment. A comparative, controlled but retrospective analysis of the results was performed of all women with infective tubal damage aged <40 years old, who had ovulatory cycles, a normal uterus and a partner with normal spermatozoa. A standardized treatment regimen was used. A maximum of three embryos were transferred. Hydrosalpinx was defined by prior hysterosalpingography and/or laparoscopy with transcervical dye injection. A total of 237 embryo transfer cycles in women with hydrosalpinges (tubal distension not visible in 151, visible but not drained in 30 and drained in 56) were compared with 705 embryo transfer cycles in women with tubal disease but no hydrosalpinx. Results were analysed in the first three cycles but also separately in the first cycle to check for bias. Success rates were higher in the first cycle, but did not significantly influence overall differences. Implantation rates were significantly reduced overall in the hydrosalpinx group (8.0 versus 13.2% for controls; P < 0.001), being 8.3% (P < 0.01) in the subgroup without evident tubal distension and 7.5% (not significant) in the drained hydrosalpinx group. This study shows that tubal damage with distal occlusion is associated with a marked reduction in embryo implantation, even in the absence of obvious fluid distension. Surgical drainage of distended hydrosalpinges appears to offer no benefit.

Adult↗

Intrauterine administration of autologous peripheral blood mononuclear cells promotes implantation rates in patients with repeated failure of IVF-embryo transfer.

BACKGROUND: There are few effective approaches to infertile patients with repeated failure in IVF-embryo transfer therapy. Since recent evidence suggests that some populations of maternal immune cells positively support embryo implantation, we have developed a new approach using peripheral blood mononuclear cells (PBMCs). METHODS: Patients who had not experienced successful pregnancy despite four or more IVF-embryo transfer sessions were enrolled in this study (n = 35, 35 cycles). PBMCs were obtained from patients on the day of oocyte retrieval and were cultured with HCG for 48 h. Two days later, PBMCs were freshly isolated from patients again, combined with cultured PBMC and then administered to the intrauterine cavity of the patients. Blastocyst transfer was performed on day 5, and the success of implantation in the PBMC-treated group was compared with that in the non-treated group. RESULTS: Clinical pregnancy rate, implantation rate and live birth rate in the PBMC-treated group (41.2, 23.4 and 35.3%; n = 17, 47 and 16, respectively) were significantly higher than those in the non-treated group (11.1, 4.1 and 5.5%; n = 18, 49 and 18, respectively). CONCLUSION: Intrauterine administration of autologous PBMC may be an effective approach to improve embryo implantation in patients with repeated IVF failures.

Adult↗

A randomized comparison of alternative methods of information provision on the acceptability of elective single embryo transfer.

BACKGROUND: The purpose of this study was to assess the effect of alternative ways of providing information about the risks of twins on couples' perceptions about elective single embryo transfer (eSET). METHODS: Couples undergoing IVF were randomized into three groups. Group 1 received a standard information pack, group 2 an extra information leaflet about twin pregnancy, and group 3 an additional discussion session. The primary outcome measure was acceptability of a hypothetical policy of eSET. Data were collected by means of a questionnaire. RESULTS: eSET was acceptable to 17 (27%), 20 (30%) and 24 (32%) couples in groups 1, 2 and 3, if it meant a slight reduction in pregnancy rates, and to 51 (82%), 55 (83%) and 53 (87%) couples, respectively, if pregnancy rates were unchanged. A fixed charge for all fresh and frozen embryo transfers following a single oocyte retrieval led to acceptability rates of 35 (57%), 36 (55%) and 38 (65%). CONCLUSIONS: Additional information, involving an extra information leaflet and face to face discussion, did not changes couples' attitudes towards eSET. Maintaining existing rates of pregnancy and offering a fixed charge for all embryo transfers resulting from an oocyte recovery may encourage more couples to consider eSET.

Embryo Transfer↗

Exogenous hormone regimens to utilize successfully mares in dioestrus (days 2-14 after ovulation) as embryo transfer recipients.

Two hormone regimens were utilized for recipient mares which were 2-14 days after ovulation at the time of non-surgical embryo transfer. In Exp. I, 20 embryos were transferred non-surgically into recipient mares which had been given 22 mg altrenogest daily starting the day of recipient ovulation. Higher (P less than 0.05) pregnancy rates (50% vs 0%) were obtained in mares which were 2-6 days after ovulation at the time of transfer compared with mares which were 7-12 days after ovulation. In Exp. II, on the day the donor mare ovulated (Day 0), 10 mg PGF-2 alpha were given to the recipient mare at 0 h, 20 mg oestradiol cypionate at 12, 24 and 36 h, 500 mg progesterone in oil at 48 h and then 22 mg altrenogest at 60, 72 and 96 h. Altrenogest (22 mg/day) was continued until Day 28 (fetal heart beat). The pregnancy rate (58% vs 10%) was higher (P less than 0.05) for the 12 recipient mares which were 10-14 days after ovulation than for the 10 mares that were 5-8 days after ovulation when treatment began. Early embryonic loss was detected in 4/8 pregnant recipient mares between Days 21 and 28 of gestation. We suggest that mares which are 2-14 days after ovulation and not in ovulation synchrony with the donor may be used as embryo transfer recipients; mares which are in early dioestrus can be given altrenogest while those in mid- to late dioestrus can be placed on the more complex hormone regimen.

Animals↗

Ovarian hyperstimulation syndrome and benign intracranial hypertension in pregnancy after in-vitro fertilization and embryo transfer: case report.

Ovarian hyperstimulation syndrome (OHSS) is a dangerous and sometimes life-threatening complication of ovulation induction with exogenous gonadotrophins. While many complications of severe OHSS are recognized we have only identified one review detailing neurological problems. This report concerns a 32-year-old patient with bilateral tubal blockage who achieved her first pregnancy following in-vitro fertilization (IVF) and embryo transfer. Shortly after embryo transfer she developed clinical signs of moderate OHSS with symptoms which were later diagnosed as benign intracranial hypertension (BIH). The BIH was treated effectively using repeated lumbar puncture and diuretics. Spontaneous labour and delivery occurred at 40 weeks' gestation. There was no neurological sequel and no recurrence of the BIH 2 years after the pregnancy. The possible link between OHSS and BIH is discussed as well as the risks of further pregnancy.

Adult↗

Investigations into the control of litter size in swine: III. A reciprocal embryo transfer study of early conceptus development.

A reciprocal embryo transfer study with Meishan and Yorkshire pigs was conducted to examine the influence of embryonic genotype and uterine environment on preimplantation embryonic growth and development. Embryos were collected from gilts on d 2 of a spontaneous estrous cycle (d 0 = onset of estrus), transferred to synchronous recipients, and collected from recipients on d 12 to obtain measurements of embryonic diameter (size) and embryonic DNA, protein, and estrogen content. No difference was detected between Meishan and Yorkshire donors for number of ovulations, number of embryos recovered, or number of cells per embryo. Embryonic genotype affected (P < .01) d-12 embryonic characteristics; Meishan embryos (n = 101) were smaller (4.7 mm diameter) and contained less DNA (4.5 micrograms) and protein (104 micrograms) than Yorkshire embryos (n = 85; 5.9 mm, 6.1 micrograms, and 149 micrograms, respectively). Embryos (n = 80) transferred into a Meishan uterus were reduced (P < .001) in diameter (4.2 vs 6.4 mm) and in DNA (3.2 vs 7.2 micrograms), protein (103.8 vs 149 micrograms), and estrogen (352 vs 1,643 pg) content compared with embryos (n = 106) transferred into a Yorkshire uterus. These data indicate that the increased prolificacy of the Meishan breed may be due to an increased embryonic survival resulting from slower growing embryos and a suppressive effect of the uterus on embryonic growth rate and estrogen secretion.

Animals↗