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State of the art in pig embryo transfer.

Embryo transfer in pigs has required surgical procedures in both donors and recipients. Over the last decade, procedures have been developed for transferring embryos by endoscopic or nonsurgical (transcervical) procedures. The feasibility of these procedures for practical application and the latest results of these new approaches are compared here. Factors affecting the current results and obstacles to be overcome in the near future are also discussed. Finally, some relevant embryo collection procedures and applications are briefly summarized.

Animals↗

Rederivation of inbred strains of mice by means of embryo transfer.

Embryo transfers were performed to rederive six inbred strains of mice, A/He, BALB/cByJ, BALB/c Lac, B10.BR/SgSnJ, C57BL/6J and DBA/2J. The aim was to determine whether it is possible to eliminate pathogens like mouse hepatitis virus (MHV) and Pasteurella pneumotropica (P.p.). The embryos were collected, handled and transferred into the oviduct of day one pseudopregnant SPF surrogate mothers under aseptic conditions. In 40.5% of the transfers, embryos developed to term. With respect to surrogate mothers delivering viable litters, 47.9% of the transferred embryos were born alive. Out of these 93.5% were reared. Virological and bacteriological examination of embryo donors verified the presence of P.p. and of antibodies against MHV in all strains. In some embryo donors P.p. could be isolated even from the uterine mucosa. However, neither in the surrogate mothers nor in the offspring could P.p. and antibodies against MHV be detected. Further bacteriological examination revealed that the offspring carried only the microbial flora received from the surrogate mother. The results indicate that embryo transfer is an appropriate tool to rederive mouse strains. In contrast to hysterectomy rederivation, embryo transfer has the advantage of avoiding postimplantational vertical transmissions of infections.

Animals↗

Incidence of microbial growth from the tip of the embryo transfer catheter after embryo transfer in relation to clinical pregnancy rate following in-vitro fertilization and embryo transfer.

A total of 110 consecutive women was studied prospectively at the time of transcervical embryo transfer following conventional in-vitro fertilization and intracytoplasmic sperm injection procedures. Microbiological cultures were performed on endocervical swabs and embryo transfer catheter tips. Positive microbial growths were observed from endocervical swabs in 78 (70.9%) women and from catheter tips in 54 (49.1%) women. The clinical pregnancy rates were 57.1% in the group of patients without growth and 29.6% in the group with positive microbial growth from catheter tips. As microbial contamination at embryo transfer may influence implantation rates, prospective studies are justified to determine whether eradication of endocervical micro-organisms is possible and whether their eradication will improve implantation rates.

Adult↗

[Effect of trial transfer on embryo transfer techniques].

OBJECTIVE: To evaluate the effect of trial transfer on embryo transfer. METHODS: Embryo transfer guided by ultrasound was performed in 114 cycles. And trial transfer was accomplished in 101 of them prior to embryo transfer. The clinical pregnancy rate and embryo implantation rate were compared between the group with trial transfer and the group without, and also among the sub-groups formed according to the difference between the trial transfer position and the actual embryo depth. RESULTS: There was no statistically significant difference in the clinical pregnancy rate and embryo implantation rate between the group with trial transfer and that without trial transfer(59.41% vs 61.54%, 35.81% vs 31.43%, P > 0.05), nor was there between the two subgroups of the group with trial transfer (61.54% vs 58.67%, 40.79% vs 34.09%, P > 0.05). CONCLUSION: As far as modern embryo transfer techniques are concerned, trial transfer can not provide accurate guidance for the depth of embryo transfer.

Adult↗

In vitro technologies related to pig embryo transfer.

Embryo transfer in swine (ETS) has been used for commercial and breeding application only to a limited extent. However this technique is an essential prerequisite for the application of new reproductive techniques in pigs. This paper will give an overview on steps of pig embryo transfer including selection and stimulation of donor sows, recovery of embryos, embryo handling and the transfer of recovered embryos into recipients. Furthermore the current status and further application of ET related in vitro technologies in pig production are described.

Animals↗

Biofluid aspects of embryo transfer.

Embryo transfer (ET) is the last stage of extracorporal fertilization during which the embryo is placed in the uterine cavity with a medium-filled catheter 2-3 days after in vitro fertilization. While fertilization in the laboratory occurs at very high rates (> 90%), the overall success of the procedure (i.e., take home baby) is still very low (< 25%) and assumed to be mainly due to implantation failure. A computational model was developed to simulate ET within the uterine cavity by a fluid-filled catheter inserted into a two-dimensional channel with oscillating walls. The results showed that the speed at which the embryos are injected from the catheter dominates the procedure and controls the velocity of their transport within the uterine cavity. ET at excessively high injection speeds may lead to ectopic pregnancies, while uterine peristalsis affects transverse dispersion only during injection at low injection speeds. The presence of the catheter within the uterus does not affect flow patterns downstream of its tip. The potential risks to implantation failure due to mechanical factors involved in the ET processes are discussed.

Animals↗

Ultrasound guidance for human embryo transfer.

Embryo transfer is facilitated with ultrasound guidance. The 16 ultrasound-guided transfers were rated easier, and there was less catheter distortion, in comparison with 12 transfers guided by "clinical feel." The benefits we ascribe to ultrasound guidance are as follows: (1) transfers can be done with the patient supine in the lithotomy position; (2) the catheter tip is accurately positioned in the fundus of the uterine cavity; (3) the ejection of the transfer bubble into the uterus and its persistence during catheter removal and documented; and (4) the continuance of the bubble is comforting to the patient. Greater experience is needed before the impact of ultrasound-guided transfer can be correlated with the rate of pregnancy continuance.

Adult↗

Development of screening systems for evaluation of materials used in mammalian embryo transfer.

Embryo transfer units use a wide variety of materials that come in contact with embryos. Studies were conducted to evaluate procedures that could be utilized to determine the toxicity of some commonly used materials in embryo collection, culture and transfer. Forty-five female mice were sacrificed on Day 3 or 4 of gestation (Day 1 = vaginal plug), and the uterus and oviducts were removed and minced. A total of 522 embryos was collected (4-cell to blastocyst stages). Four to 16 cell embryos were cultured in Phosphate Buffered Saline (PBS) plus 20% fetal bovine serum. Morula to blastocyst stage embryos were cultured in Nutrient Mixture F10 (HAM) plus 20% fetal bovine serum gassed with 5% CO(2), 5% O(2) and 90% N(2). In Experiment I, embryos and culture media were placed in a covered embryological watch glass (EWG, control) or sealed in the lumen of a siliconized Foley catheter or a section of 1) latex tubing, 2) tygon tubing or 3) silastic tubing. In Experiment II, embryos were placed in EWG and cultured alone (control) or cocultured with sections of 1) tygon tubing, 2) silastic tubing or 3) latex tubing. In Experiment III, embryos were cultured in covered plastic petri dishes containing 15 ml of media, alone (control) or co-cultured with two new plunger tips from sterile Monoject syringes. All embryos were cultured at 32 to 34 degrees C for 24 h. The Criterion used for development was two or more cellular divisions within the 24-h period. Embryo development in Experiment I was lower (P<0.05) in latex (0%) and tygon (24%) tubing and in the siliconized Foley catheter (2%) than in silastic tubing (51%) and the EWG (46%), which did not differ. Experiment II embryos that were co-cultured with latex tubing (5%) showed very little development as compared with those co-cultured with tygon tubing (76%), silastic tubing (76%) and EWG (93%), the last of which were not significantly different. Embryos co-cultured with Monoject syringe plunger tips had a reduced embryo development rate compared to embryos in the control group (0% vs 52%). Although the embryos did not remain in contact with these seemingly toxic materials for prolonged periods, our results indicate that a significant reduction in embryo viability may occur due to this exposure.

Journal Article↗

Effect of the number and depth of embryos transferred and unilateral or bilateral transfer in tubal embryo transfer (TET).

PURPOSE: Our purpose was to evaluate the possible effects of the number of embryos transferred, the depth of embryos placed within the tube(s), and unilateral or bilateral tubal transfer on pregnancy initiation in tubal embryo transfer (TET). METHODS: One hundred eight consecutive TET cycles were analyzed. Oocyte retrievals were carried out by transvaginal ultrasound-guided aspiration of follicles. Forty-eight hours after oocyte retrieval, the developing embryos at the stage of two to four cells were transferred into the fallopian tube(s) by laparoscopy. A maximum of four embryos was transferred to each patient. RESULTS: The pregnancy rates were similar among the cycles in which two, three, or four embryos were transferred. In addition, there was no significant difference in the pregnancy rate whether the embryos were deposited > 4 cm or between 3 and 4 cm into the tube(s). Although the pregnancy rate was greater in cycles of bilateral tubal transfer, the difference from that of unilateral transfers was not significant. CONCLUSION: Our data indicate that when two to four embryos were transferred and the embryos were placed > or = 3 cm within the tube(s), unilateral or bilateral tubal transfer had little influence on the ultimate success of TET.

Embryo Transfer↗

Comparison of concurrent pregnancy rates for in-vitro fertilization--embryo transfer, pronuclear stage embryo transfer and gamete intra-fallopian transfer.

Concurrent pregnancy and implantation (sacs/embryos transferred) rates were compared for 84, 77 and 49 cases of in-vitro fertilization-embryo transfer (IVF-ET), pronuclear stage embryo transfer (PROST) and gamete intra-Fallopian transfer (GIFT), respectively. All cases reported occurred during an 18-month interval since the initiation of PROST by our programme. Leuprolide acetate was used with follicle stimulating hormone and human menopausal gonadotrophin for follicular stimulation of all but donor oocyte cases (n = 9). Clinical pregnancy (per transfer) and implantation rates were significantly higher (P less than 0.03) for PROST (52.4%, 20.2%) in comparison with IVF-ET (26.9%, 11.4%). Rates for GIFT (48.9%, 18.4%) were not significantly higher (P = 0.10, 0.14) than for IVF-ET. This was probably due to the lower number of GIFT than PROST procedures performed. The total pregnancy rate for GIFT (biochemical, ectopic and clinical combined) was significantly greater (P less than 0.05) than for IVF-ET. Pregnancy and implantation rates for PROST and GIFT were similar. These results support the use of PROST rather than IVF-ET for all cases in which the woman has one functional Fallopian tube. Furthermore, to maintain equivalent rates of pregnancy with PROST and GIFT, it is suggested that GIFT should not be used for cases of male-factor infertility without first documenting normal rates of in-vitro fertilization with PROST.

Adult↗

Uterine position at real embryo transfer compared with mock embryo transfer.

BACKGROUND: The purpose of this study was to determine the consistency in the uterine position between mock and real embryo transfer. METHODS: We reviewed 996 consecutive embryo transfer cycles (585 patients); 74% of patients had an anteverted (AV) uterus and 26% had a retroverted (RV) uterus at mock embryo transfer. All mock and real embryo transfers were performed under abdominal ultrasound guidance. RESULTS: Of 623 fresh embryo transfers in patients with an AV uterus at mock embryo transfer, only 2% became RV, while 55% of 213 embryo transfers in patients with an RV uterus on mock embryo transfer converted to AV at real embryo transfer (P < 0.0001). For frozen-thawed embryo transfer, 12% of AV uteri at mock embryo transfer became RV, while 33% of RV uteri became AV (P = 0.01). CONCLUSIONS: Our data suggest that an RV uterus at mock embryo transfer will often change position at real embryo transfer. Misdirecting the embryo transfer catheter can be avoided by accurate knowledge of the uterine position at the time of embryo transfer, which can be more accurately assessed by routine ultrasound guidance. Additionally, patients with an RV uterus at mock embryo transfer should still present with a full bladder for embryo transfer, since a significant number will convert to an AV position.

Cryopreservation↗

Statistical models predicting embryo survival to term in cattle after embryo transfer.

Embryo survival to term in recipient cattle is highly variable. We examined calving data in the published literature to determine whether a model of binomial independence or a model which includes an embryo (e) and recipient term (r), adequately explain observed embryo survival rates following attempts to induce twin calving using transfer of two embryos. To achieve this we examined 32 published papers which provided us with 47 sets of data concerning 4560 recipients with either 0, 1 or 2 calves born. In each set of data, the observed embryo survival rate to term (p) (number of calves born/number of embryos) was calculated and the expected number of recipients with either 0, 1 or 2 calves born was determined, assuming a binomial distribution. Parameters for the second model were estimated using maximum-likelihood procedures. The model of embryo independence was rejected in 85% of the sets of data, suggesting that factors other than the embryo are important sources of variation in embryo survival or loss. The proposed e and r model of embryo survival adequately describes the published data in recipients receiving either single or twin embryos. In general, only 50-70% of embryos and recipients are sufficiently competent to result in a calving. Variation among laboratories producing either in vitro or in vivo derived embryos was due to variation in recipient and not embryo competence. It is argued that e rather than observed embryo survival rate, and r rather than observed pregnancy rate, should be used to compare differences among embryo treatments and groups of recipients, respectively. Acceptance of this proposition should permit faster progress in identifying the biology of superior embryos and recipients, which is a prerequisite to improving embryo survival rate in cattle. Collectively, the published data are not consistent with a model of embryo independence, and that a model of embryo survival to term which recognises recipient as well as embryo contributions to embryo survival may be more appropriate in cattle.

Animals↗

[Differential indications for surgical technics in reproductive medicine--microsurgery, in vitro fertilization and embryo transfer, gamete intrafallopian transfer and tubal embryo transfer].

The birth of the first baby following in-vitro fertilisation and embryo transfer (IVF/ET) in 1978 and the introduction of gamete intrafallopian transfer (GIFT) in 1984 have increased the treatment modalities in operative reproductive medicine. In tubal pathology, there are besides micro-surgery now so-called additive methods available for treating infertility. With regard to the indications, there has been severe confusion, and it is therefore imperative to define special indications for IVF/ET, microsurgery and GIFT. We do not consider these treatment modalities as alternative methods. Reproductive centres should offer all these treatments to guarantee an appropriate individual programme for each couple. Therefore, knowledge of pregnancy rates is a prerequisite for the doctor. The aim of this paper is, to define the optimal therapy while taking into account the individual problems of each couple.

Embryo Transfer↗

Anti-leucocyte antibodies and embryonic mortality in embryo transferred cows.

Cows carrying unrelated transferred embryos (ET) produced anti-leucocyte serum antibodies (aLA) more often than cows carrying their own embryos. Cows carrying the ET showed a higher frequency of cytotoxic reactions against leucocytes from 40-60 randomly chosen cows than individuals carrying their own embryos. The percentage of animals with aLA was higher in cows carrying their second or third transferred embryo than in those with their first transferred embryo. There was no change in the frequency of cytotoxic reactions with repeated pregnancies from transferred embryos. There was no difference in the toxicity of aLA in normal pregnant cows and those carrying transferred embryos. Embryonic mortality (EM) of 35, 73 and 88% was noted during pregnancies from the first, second and third successful ET, respectively. Mortality of 48% occurred in the first pregnancy following an unsuccessful ET. Embryonic mortality of 31% occurred in cows simultaneously carrying their own and a transferred embryo. A direct relationship between the presence of aLA and EM in recipients was not proved. Other fertility problems may lead to EM in cows subjected to repeated transfer of foreign embryos.

Abortion, Veterinary↗

Comparison of day 2 tubal embryo transfer with day 3 embryo transfer into the uterus of intracytoplasmic sperm injection derived human embryos.

OBJECTIVE: To compare the results of embryo transfer into the uterus (ET) performed on day 3 vs. tubal embryo transfer (TET) on day 2 in couples diagnosed with male factor infertility. MATERIALS AND METHODS: We collected data from 34 and 110 patients who underwent intracytoplasmic sperm injection (ICSI) followed by either TET on day 2 or ET on day 3 (January 2001-June 2005), respectively. All couples were diagnosed with male factor infertility; there were no tubal or uterine factors, diminished ovarian reserve, pelvic adhesions, history of polycystic ovary syndrome or previous ovarian surgery. RESULTS: The clinical pregnancy rates of day 2 TET and day 3 ET following ICSI were 64.71% and 57.27% (p > 0.05), respectively. The corresponding implantation rates were 35.47% and 29.58% (p > 0.05), respectively. CONCLUSION: The implantation and clinical pregnancy rates of ET on the 3rd day following ICSI were similar to the rates obtained with TET 2 days after ICSI. Therefore, ET performed 3 days after ICSI should be the first choice for couples with male factor infertility, while day 2 TET remains a good alternative, especially for women with normal tubal function and known difficulties of transcervical ET.

Embryo Transfer↗

A real-life prospective health economic study of elective single embryo transfer versus two-embryo transfer in first IVF/ICSI cycles.

BACKGROUND: We analysed the difference in maternal, neonatal and total costs after single (SET) versus double day 3 embryo transfer (DET). METHODS: We performed a two-centre prospective study of women in their first IVF/ICSI cycle choosing between SET or DET. Infertility treatment data were gathered from a database; maternal and neonatal outcome data from a case report form (CRF); health economic data from medical acts registered in the CRF for the outpatient part and from hospital bills. SET was performed in 206/367 (56.1%) and DET in 161/367 (43.9%) women. RESULTS: In all, 367 transfers yielded 186 positive pregnancy tests, 148 ongoing pregnancies and 136 live deliveries (50.7, 40.3 and 37.1% per embryo transfer) of which 15 (11.0%) were twins. Live birth rate was 37.4% for SET, 36.6% for DET. Intention-to-treat analysis showed differences for: duration of pregnancy (SET: 39.0 +/- 1.4 versus DET: 38.3 +/- 2.2 weeks; P = 0.055), percentage prematurity (8.5 versus 23.8%; P = 0.033), percentage of neonates hospitalized (5.7 versus 17.9%; P = 0.121) and duration of neonatal hospitalization (6.3 +/- 2.2 versus 10.3 +/- 10.1 days; P = 0.01). Total cost after DET was higher (SET: 4700 +/- 3239 versus DET: 8613 +/- 10 105; P = 0.105), due to significantly higher neonatal costs (451 +/- 957 versus 3453 +/- 8154; P < 0.001) and not to differences in maternal costs (4250 +/- 2882 versus 5160 +/- 4106; P = 0.152). CONCLUSIONS: This prospective health economic study shows that transfer of a single top quality embryo is equally effective as, but substantially cheaper than, double embryo transfer in women <38 years of age in their first IVF/ICSI cycle.

Birth Rate↗

Impact of patients' choice for single embryo transfer of a top quality embryo versus double embryo transfer in the first IVF/ICSI cycle.

BACKGROUND: The aim of this study was to evaluate the impact of transferring a single top quality embryo in the first IVF/ICSI cycle of patients <38 years old who chose to have one or two embryos transferred. METHODS: A total of 262 patients participated in the study, and 243 transfers were performed: 156 (64%) patients chose the transfer of a single top quality embryo, if available, and two non-top quality embryos if not available; 87 (36%) patients chose to have a double embryo transfer regardless of embryo quality. RESULTS: In the first group an ongoing pregnancy rate of 40% (63/156) with a twin pregnancy rate of 2% (1/63) was achieved. In the second group the ongoing pregnancy rate was 44% (38/87) with 26% (10/38) twin pregnancies. In the patient group with only one embryo transferred, irrespective of the patient's choice, the ongoing pregnancy rate was 43% (54/127) with no twin pregnancies. For the study population as a whole, the ongoing pregnancy rate was 42% (101/243) with 11% (11/101) twins. CONCLUSION: We conclude that the introduction of single embryo transfer in the first IVF/ICSI cycle is highly acceptable in women <38 years old.

Adult↗