scholarly journals Coculture of Human Embryos with Autologous Human Endometrial Epithelial Cells in Patients with Implantation Failure1

1999 ◽  
Vol 84 (8) ◽  
pp. 2638-2646
Author(s):  
Carlos Simón ◽  
Amparo Mercader ◽  
Juan Garcia-Velasco ◽  
George Nikas ◽  
Carlos Moreno ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Embryo Transfer ◽  
Cardiac Activity ◽  
Oocyte Donation ◽  
Pregnancy Rates ◽  
Blastocyst Transfer ◽  
Human Embryos ◽  
Blastocyst Formation ◽  
Ovum Donation ◽  
Endometrial Epithelial Cells

We have developed a coculture system with autologous human endometrial epithelial cells (AEEC) that retained many features of human endometrial epithelium. Implantation failure (IF; >3 previous cycles failed with 3–4 good quality embryos transferred) is a distressing condition in which 2-day embryo transfer repetition is the routine option. The objective of this study was to investigate the basics and to evaluate prospectively the clinical value of embryo coculture on AEEC and blastocyst transfer with their own oocytes [in vitro fertilization (IVF) patients] or with donated oocytes (oocyte donation patients) compared to a routine day 2 embryo transfer for patients with IF. Scanning electron microscopy and mouse embryo assays demonstrate that EEC from fertile and IF patients were morphologically and functionally similar; similar findings were observed in EEC obtained from fresh or frozen endometria. Clinically, 168 IVF cycles were performed in 127 patients with 3.8 ± 0.2 previously failed cycles, and 80 cycles were performed in 57 patients undergoing oocyte donation with 3.0 ± 0.2 previously failed cycles. Twenty IVF patients and 15 ovum donation patients with 3 previously failed cycles in whom a 2-day embryo transfer was performed were used as controls. In 88% of ovum donation cycles, at least 2 blastocysts were available for transfer, with 60.1% blastocyst formation; 2.2 ± 0.1 blastocysts were transferred/cycle, and 36 pregnancies (determined by fetal cardiac activity) were obtained (32.7% implantation and 54.5% pregnancy rates). In 168 IVF cycles, 8.1 ± 0.2 embryos/cycle started coculture, resulting in 49.2% blastocyst formation; 2.3 ± 0.2 blastocysts were transferred/cycle, and 29 clinical pregnancies were obtained (11.8% implantation and 20.2% pregnancy rates). Fifteen cycles were canceled (9%). In oocyte donation patients with IF undergoing 2-day embryo transfer, implantation and pregnancy rates were significantly lower (4.5% and 13.3%; P < 0.01) than with coculture; however, in IVF patients with IF, results with day 2 transfer (10.7% and 35%) were similar to those with coculture. The present study demonstrates that coculture of human embryos with AEEC and blastocyst transfer is safe, ethical, and effective and constitutes a new approach to improve implantation in patients with IF undergoing ovum donation, but not in IVF patients.

scholarly journals The effects of hyaluronate-containing medium on human embryo attachment to endometrial epithelial cells in vitro

2020 ◽  
Vol 2020 (2) ◽  
Author(s):  
Peter T Ruane ◽  
Chelsea J Buck ◽  
Phoebe A Babbington ◽  
Wedad Aboussahoud ◽  
Stéphane C Berneau ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Embryo Transfer ◽  
Human Embryo ◽  
Research Network ◽  
Human Embryos ◽  
Ishikawa Cell ◽  
Practitioner Training ◽  
Cell Layers ◽  
Endometrial Epithelial Cells

Abstract STUDY QUESTION Does embryo transfer medium containing hyaluronate (HA) promote the attachment phase of human embryo implantation? SUMMARY ANSWER HA-containing medium does not promote human blastocyst attachment to endometrial epithelial cells in vitro. WHAT IS KNOWN ALREADY Embryo transfer media containing high concentrations of HA are being used to increase implantation and live birth rates in IVF treatment, although the mechanism of action is unknown. STUDY DESIGN, SIZE, DURATION Expression of HA-interacting genes in frozen-thawed oocytes/embryos was assessed by microarray analysis (n = 21). Fresh and frozen human blastocysts (n = 98) were co-cultured with human endometrial epithelial Ishikawa cell layers. Blastocyst attachment and the effects of a widely used HA-containing medium were measured. PARTICIPANTS/MATERIALS, SETTING, METHODS Human embryos surplus to treatment requirements were donated with informed consent from several ART centres. Blastocyst-stage embryos were transferred at day 6 to confluent Ishikawa cell layers; some blastocysts were artificially hatched. Blastocyst attachment was monitored from 1 to 48 h, and the effects of blastocyst pre-treatment for 10 min with HA-containing medium were determined. MAIN RESULTS AND THE ROLE OF CHANCE Human embryos expressed the HA receptor genes CD44 and HMMR, hyaluronan synthase genes HAS1–3, and hyaluronidase genes HYAL1–3, at all stages of preimplantation development. Attachment of partially hatched blastocysts to Ishikawa cells at 24 and 48 h was related to trophectoderm grade (P = 0.0004 and 0.007, respectively, n = 34). Blastocysts of varying clinical grades that had been artificially hatched were all attached within 48 h (n = 21). Treatment of artificially hatched blastocysts with HA-containing medium did not significantly affect attachment at early (1–6 h) or late (24 and 48 h) time points, compared with control blastocysts (n = 43). LIMITATIONS, REASONS FOR CAUTION Using an adenocarcinoma-derived cell line to model embryo-endometrium attachment may not fully recapitulate in vivo interactions. The high levels of blastocyst attachment seen with this in vitro model may limit the sensitivity with which the effects of HA can be observed. WIDER IMPLICATIONS OF THE FINDINGS Morphological trophectoderm grade can be correlated with blastocyst attachment in vitro. HA-containing medium may increase pregnancy rates by mechanisms other than promoting blastocyst attachment to endometrium. STUDY FUNDING/COMPETING INTEREST(S) This work was funded by a grant from the Wellbeing of Women, the NIHR Local Comprehensive Research Network and NIHR Manchester Clinical Research Facility, the Department of Health Scientist Practitioner Training Scheme, and the Ministry of Higher Education, The State of Libya. None of the authors has any conflict of interest to declare.

scholarly journals Cleavage Stage versus Blastocyst Stage Embryo Transfer in Oocyte Donation Cycles

Medicina ◽  
2019 ◽  
Vol 55 (6) ◽  
pp. 293 ◽  
Author(s):  
Kontopoulos ◽  
Simopoulou ◽  
Zervomanolakis ◽  
Prokopakis ◽  
Dimitropoulos ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Oocyte Donation ◽  
Blastocyst Stage ◽  
Pregnancy Rates ◽  
Clinical Pregnancy ◽  
Cleavage Stage ◽  
Sonographic Guidance ◽  
Stage Embryo ◽  
Day 3 Embryo ◽  
Cryopreserved Embryos

Background and Objective: During the last few years, a trend has been noted towards embryos being transferred at the blastocyst stage, which has been associated with improved rates regarding implantation and clinical pregnancy in comparison to cleavage stage embryo transfers. There is a limited number of studies investigating this notion in oocyte donation cycles employing cryopreserved embryos. The aim of this study is to evaluate the implantation potential and clinical pregnancy rates between the day 3 cleavage stage and blastocyst stage embryo transfers in oocyte donation cycles employing vitrified embryos. Methods: This is a retrospective evaluation of oocyte donation frozen–thawed transfers completed in our clinic from January 2017 to December 2017. Intracytoplasmic sperm injection was conducted for all oocytes. Following fertilization, all embryos were cryopreserved either at the cleavage or blastocyst stage. Embryo transfer of two embryos was performed under direct sonographic guidance in all cases. Results: Our results confirmed a 55.6% clinical pregnancy (CP) resulting from day 3 embryo transfers, a 68.8% CP from day 5, and 71.4% CP from day 6. Significantly improved pregnancy rates were related to embryo transfers at the blastocyst stage when compared to cleavage stage transfers (68.9% and 55.6% respectively, p = 0.016). The risk with regards to multiple pregnancies was similar. Conclusion: Our findings indicate that in oocyte donation cycles employing vitrified embryos, embryo transfer at the blastocyst stage is accompanied with a significant improvement in pregnancy rates and merits further investigation.

scholarly journals Evaluation of Pregnancy Outcomes of Vitrified-Warmed Blastocyst Transfer before and after Endometrial Receptivity Analysis in Identical Patients with Recurrent Implantation Failure

2021 ◽  
pp. 1-7
Author(s):  
Yuta Kasahara ◽  
Tomoko Hashimoto ◽  
Ryo Yokomizo ◽  
Yuya Takeshige ◽  
Koki Yoshinaga ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Embryo Transfer ◽  
Pregnancy Outcomes ◽  
Endometrial Receptivity ◽  
Pregnancy Rates ◽  
Blastocyst Transfer ◽  
Implantation Failure ◽  
Recurrent Implantation Failure ◽  
Clinical Value ◽  
Ongoing Pregnancy Rate

Background:The clinical value of personalized embryo transfer (pET) guided by the endometrial receptivity analysis (ERA) tests for recurrent implantation failure (RIF) cases is still unclear. The aim of this study is to clarify the efficacy of ERA leading to personalization of the day of embryo transfer (ET) in RIF patients. Methods: A retrospective study was performed for 94 patients with RIF who underwent ERA between July 2015 and December 2019. Pregnancy outcomes in a previous vitrified-warmed blastocyst transfer (previous VBT) and a personalized vitrified-warmed blastocyst transfer (pVBT) in identical patients were compared. The details of each pVBT were further analyzed between patients in a non-displaced group, which indicated “receptive” cases in ERA results and those who were in the displaced group, which indicated “non-receptive” cases. Results:When the pregnancy rate, both per patient and per transfer cycle, of previous VBT and pVBT were compared, a significant increase in pVBT was observed between the two methods (5.3% vs. 62.8%, 4.4% vs. 47.9%, respectively). The pregnancy rates, implantation rates, and clinical pregnancy rates of the first pVBT were significantly higher in the displaced group than the non-displaced group. The cumulative ongoing pregnancy rate of the displaced group tended to be higher compared to that of the non-displaced group in the first pVBT, although the difference was not statistically significant (51.0% vs. 31.1%, [Formula: see text] = 0.06). Conclusions:Our study demonstrates that pVBT guided by ERA tests may improve pregnancy outcomes in RIF patients whose window of implantation (WOI) is displaced, and its effect may be more pronounced at the first pVBT. The displacement of WOI may be considered to be one of the causes of RIF, and its adjustment may contribute to the improvement of pregnancy outcomes in RIF patients.

P–354 Analysis of pregnancy and miscarriage rates in anti-centromere antibodies (ACA)-positive patients treated with ART

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Y Kida ◽  
M Tokoro ◽  
H Kitasaka ◽  
T Yoshimura ◽  
N Fukunaga ◽  
...  
Keyword(s):  
Embryo Transfer ◽  
Antinuclear Antibody ◽  
Assisted Reproductive Technologies ◽  
Formation Rate ◽  
Reproductive Technologies ◽  
Pregnancy Rates ◽  
Human Embryos ◽  
Antibody Testing ◽  
Miscarriage Rate ◽  
Significant Difference

Abstract Study question Do ACA have an effect on pregnancy and miscarriage rates of human embryos? Summary answer The present results suggest that in ACA-positive cases, the pregnancy rate per transfer was significantly lower, although the miscarriage rate was not affected. What is known already We have previously shown that patients with high levels of anti-centromere antibody (ACA), (one of the anti-nuclear antibodies (ANA)), frequently have dispersal of the female chromosomes in the cytoplasm. Additionally, we reported that the clinical outcome was characterized by a low oocyte maturation rate following ovum pick up and high multiple pronuclear formation rate after fertilization. However, the post-implantation course of embryos with ACA-positive cases has not yet been reported. Therefore, in this study, we analyzed the pregnancy and miscarriage rates in ACA-positive patients treated with Assisted Reproductive Technologies (ART). Study design, size, duration 6581 patients who underwent embryo transfer after antinuclear antibody testing between January 2014 and February 2020 were included in the analysis. Participants/materials, setting, methods The subjects were classified into three groups: ANA-negative (without ACA or any other ANA), ACA-positive (with only ACA) and ANA-positive (with ANA but not ACA). The cycle in which the gestational sac was confirmed was considered a positive pregnancy. The pregnancy and miscarriage rates were compared among the groups using “Ryan Test” for statistical analysis. Main results and the role of chance Of the 6581 eligible cases, the incidence of antinuclear antibody were 71.3% (4695/6581; ANA-negative), 0.9% (61/6581; ACA-positive) and 27.7% (1825/6581; ANA-positive). The pregnancy rates based on the total number of embryo transfer cycles for each were ANA-negative: 31.5% (5283/16792), ACA-positive: 17.6% (41/233), and ANA-positive: 32.4% (1891/5833). The pregnancy rates were significantly lower in the ACA-positive group than in the other groups. The miscarriage rate was 29.4% (1553/5283) in ANA-negative, 31.7% (13/41) in ACA-positive, and 28.0% (529/1891) in ANA-positive, with no significant difference between the three groups. Limitations, reasons for caution Retrospective analysis Wider implications of the findings: ACA-positive patients may benefit from a treatment strategy to increase the absolute number of oocytes by obtained in order to increase the chances of normal fertilization and attainment of implantation. Trial registration number none

scholarly journals Embryonic Regulation of Integrins β3,α 4, and α1 in Human Endometrial Epithelial Cells in Vitro1

1997 ◽  
Vol 82 (8) ◽  
pp. 2607-2616 ◽  
Author(s):  
Carlos Simón ◽  
MarÍa José Gimeno ◽  
Amparo Mercader ◽  
José Enrique O’Connor ◽  
José RemohÍ ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Epithelial Cells ◽  
Blastocyst Stage ◽  
Human Embryos ◽  
Adhesion Assay ◽  
Fertilization Program ◽  
Human Blastocyst ◽  
Embryonic Regulation ◽  
Endometrial Epithelial Cells

In the present study, we examined the embryonic regulation ofβ 3 integrin in human endometrial epithelial cells (EEC) at the protein level and analyzed putative embryonic factors responsible for this regulation. The model employed is based on a clinical in vitro fertilization program in which single human embryos were cocultured with EEC until blastocyst stage and then transferred back to the uterus. After embryo transfer, EEC wells were divided according to the embryonic status reached: EEC with embryos that achieved the blastocyst stage, EEC with arrested embryos, and EEC without embryos. Immunostaining for β3 was positive in plasma membrane of EEC. Flow cytometry showed a mean percentage ofβ 3-stained cells of 24.1 ± 5.7 in EEC cocultured with embryos that achieved the blastocyst stage (n = 13) vs. 9.5 ± 1.6 (P < 0.05) in those EEC cultured with arrested embryos (n = 12). Immunostaining for α1 and α4 integrins was negative in EEC monolayers studied, regardless of the presence or absence of embryos, and these findings were confirmed by flow cytometry. The possibility that the embryonic IL-1 system and leukemia inhibitory factor were involved in the endometrial β3 up-regulation was investigated by neutralizing experiments demonstrating a significant inhibition of β3-stained cells when EEC monolayers were cultured in the presence of EEC/blastocyst-conditioned media with (n = 4) vs. without (n = 8) antihuman interleukin (IL)-1α + IL-1β (1.65% vs. 14.6%; P < 0.05). Dose-response experiments further demonstrated an up-regulation of β3 positive cells when IL-1α + IL-1β were added to the medium at a concentration of 10 pg/mL compared with control medium without added cytokines (40% vs. 20%, n = 4). The functional relevance of the EEC β3 up-regulation was tested using a mouse blastocyst adhesion assay. More mouse blastocysts attached to EEC previously in contact with human blastocyst (72.7%) compared with those EEC previously in contact with arrested embryos (40%). Our results demonstrate the selective effect of a developing human embryo on EEC expression of β3, which is maximal when a human blastocyst instead of an arrested embryo is considered. Furthermore, the embryonic IL-1 system seems to be involved in the EECβ 3 up-regulation, reinforcing the concept of precise paracrine cross-talk between blastocyst and endometrial epithelium during embryonic implantation.

scholarly journals Endometrial Vascularity: Its Relation to Implantation Rates

2012 ◽  
Vol 3 (2) ◽  
pp. 48-50 ◽  
Author(s):  
Sonal Panchal ◽  
Chaitanya Nagori
Keyword(s):  
Embryo Transfer ◽  
Endometrial Thickness ◽  
Pregnancy Rates ◽  
Junctional Zone ◽  
Ovum Donation ◽  
Biochemical Pregnancy ◽  
Implantation Rates ◽  
Β Hcg ◽  
Endometrial Vascularity

ABSTRACT Aim The aim of this study was to find out if endometrial vascularity can be used as a predictive factor for implantation Materials and methods This is a retrospective study of 500 ovum donation—embryo transfer cycles, with basal S FSH > 25. Those with endometrial thickness of >8 mm with intact junctional zone and uterine artery PI < 3.2 were taken for embryo-transfer. Vaginal micronized progesterone was started from the day of ovum pick up of the ovum donor. Two fresh grade 1, 4-6 cell embryos, were transferred on day 3. Progesterone support was continued till the day of β-hCG. β-hCG was checked in all patients followed by USG 2 weeks later. Results were observed for four groups, depending on vascularity zones 1, 2, 3, 4.2 Follow-up with ultrasound was done till 8 weeks for ongoing pregnancy. Results The biochemical pregnancy rates and ultrasound evidenced pregnancy rates were very high when vascularity was seen in zone 3 and 4 of endometrium with low abortion rates. Conclusion We believe that endometrial vascularity is an important parameter to assess the implantation potential of the endometrium. How to cite this article Nagori C, Panchal S. Endometrial Vascularity: Its Relation to Implantation Rates. Int J Infertility Fetal Med 2012;3(2):48-50.

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