The effect of oocyte vitrification on subsequent embryo developmental morphokinetics and blastocyst formation: a prospective time-lapse sibling oocyte study

2018 ◽  
Vol 37 ◽  
pp. e12-e13
Author(s):  
Stefanie De Gheselle ◽  
Vanessa Muyshond ◽  
Kelly Tilleman ◽  
Petra De Sutter
2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
K Malhotra ◽  
J Malhotra ◽  
N Malhotra ◽  
N Malhotra

Abstract Study question Do embryologists need additional training or certifications before using add on techniques in the lab ? Summary answer Out of 173 respondents majority feel add on techniques require training and/or certifications, the mode of training varies for different add ons. What is known already Cochrane reviews have suggested that minimal evidence exists for the use of add on treatments in ART, the data on the prevalence of add ons in IVF is unclear but the presence of technologies in ART laboratories world over suggests a increasing trend of adoption of unproven techniques. No data exists suggesting the role of embryologists in performing the Add on techniques and how their training or lack of, can impact patient safety. The most common method of training comes from the manufacturers and there is a lack of structured trainings for add on treatments worldover. Study design, size, duration An internet based survey was designed keeping in mind commonly available laboratory add ons. It comprised of 9 sections and a total of 18 multiple choice questions. Answer choices ranged from a simple yes or no to more complex choices suggesting the type of training and the potential benefits of training. Participants/materials, setting, methods The Survey includes results from 173 embryologists from india with varying degree of experience. Add ons included in the survey were Sperm DNA Fragmentation test, IMSI, PICSI, Microfluidics, MACS, Advanced culture media, Oocyte vitrification, Assisted hatching, Time Lapse imaging, spindle view and Electronic Witnessing. The most common practice suggestions were tabulated and identified. Main results and the role of chance The survey reports huge need for training for different add on treatments (SDF –91.4%, IMSI - 81.2%, PICSI- 66.5%, Microfluidics- 55.9%, MACS –55.3%, MicroTese- 86.9%, using advanced culture medias{Calcium ionophore- 73.4%, Hyaluronan rich media- 52.1%, growth cytokine rich media–48.5%, Theophylline for sperm motility–50.9%},oocyte Vitrification 85.5%, Assisted Hatching 75.4%, Time-lapse and Electronic witnessing 77.3%, Polarised microscopy for spindle assessment 73.5%). The Most preferred mode of training for more invasive procedures was Hands on training, followed by On the job training and validation followed by workshops(SDF- 62.6%, IMSI- 61%,PICSI–56.1%, MACS and Microfluidics 38.8%,microTese- 50.6%,Oocyte freezing 85.5%, assisted hatching 67.8%,Time-lapse and electronic witnessing 77.3%, Spindle view 73.5% ). The most preferred mode of training for non invasive procedures was Workshops and Observerships, followed by CME’s, followed by product Leaflets(44.4%,42.1%,13.5% respectively). The most common answer for the disadvantages of not being trained was unable to use the technology to its fullest potential(88.5%), whereas the most common answer for the benefit of being trained was better outcomes with said technologies(76.6%). Limitations, reasons for caution This study includes responses from embryologists who have varied levels of experience, while the need for training can be established based on these results, a junior level embryologist might answer the survey differently as compared to a senior or a lab director. Wider implications of the findings: This is a first of its kind large survey, suggesting the need for training and validation from the perspective of the embryologist. This data can be used in formulating guidelines for future trainings and can help regulators in deciding on the most preferred mode of training. Trial registration number -


2021 ◽  
Vol 159 ◽  
pp. 147-152
Author(s):  
Karolina Fryc ◽  
Agnieszka Nowak ◽  
Barbara Kij ◽  
Joanna Kochan ◽  
Pawel M. Bartlewski ◽  
...  

2017 ◽  
Vol 108 (3) ◽  
pp. e60
Author(s):  
M. Arvizu ◽  
I. Dimitriadis ◽  
G. Christou ◽  
C. Tanrikut ◽  
C.L. Bormann ◽  
...  

2014 ◽  
Vol 28 ◽  
pp. S3
Author(s):  
Sandrine Chamayou ◽  
Carmelita Alecci ◽  
Giorgia Storaci ◽  
Carmen Claudia Ragolia ◽  
Antonino Guglielmino

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M I Papadopoulou ◽  
A Papatheodorou ◽  
A Vorniotaki ◽  
N Christoforidis ◽  
A Chatziparasidou

Abstract Study question Is multinucleation during cleavage stage correlated with the ploidy status of embryos and how does it affect the clinical outcome? Summary answer The presence of multinucleated embryos does not affect clinical outcome, although the risk of aneuploidy is higher in multinucleated embryos. What is known already Multinucleated blastomeres (ΜΝ) of cleavage stage embryos has been reported widely in scientific literature. Multinucleation has been associated with diminished embryo developmental competency and clinical outcomes such as lower implantation. Although this is an intriguing subject of research, it is not clear yet whether multinucleation is related to aneuploidy. Morphological irregularities such as multinucleation in blastomeres became a constant finding only after the perpetually evolving technology of time-lapse culture of embryos which in combination with PGT-A analysis, creates new research paths which aim to develop a new tool for selection or deselection of embryos for transfer. Study design, size, duration This retrospective study, included 97 PGT-A cycles, performed at Embryolab fertility clinic from May 2017 to December 2020, all cultured in time-lapse incubator (EmbryoScope). Two study groups were formed; the MN Group consisted of PGT-A cycles with at least one multinucleated embryo (n = 56) and the Control Group in which all PGT-A cycles had no multinucleated embryos (n = 38). Euploidy rate, type of chromosomal abnormality, cumulative pregnancy and live birth rates were compared between the two groups. Participants/materials, setting, methods Embryos were annotated for the existence of multinucleated blastomeres on Day 2 of their development. Biopsy was performed on Days 5/6 and embryos were genetically tested. One or two euploid embryos were transferred. Euploidy rate and clinical outcomes between the two groups were compared. Within the MN group, euploidy rate between multinucleated and non-multinucleated embryos was compared. For abnormal embryos, association of multinucleation with the type of abnormality was tested. SAS statistical analysis was performed. Main results and the role of chance Mean female age was 35.93 years in the MN group and 38.39 years in the control group. Blastocyst formation rate (expressed per fertilised oocytes) was similar between MN and Control group (74% vs 76%, p = 0.6303). In the MN group, 56 cases resulted in 44 embryo transfers while in the control group 38 cases resulted in 23 embryo transfers. Pregnancy rates (59.09% vs 65.21%, p = 0.6255) and clinical pregnancy rates (45.45% vs 39.13%, p = 0.4245) were not significantly different between MN and Control group. Initially, cumulative live birth rate was found to be significantly higher in the MN group compared to the Control group (62.96% vs 33.34%, p = 0.0417). However, when logistic and poisson regression was applied, it became obvious that this difference was not affected by multinucleation but from other factors such as female age. When comparing multinucleated and non-multinucleated embryos within the MN group, it was found that the mean number of euploid embryos was significantly higher in the non-multinucleated subgroup of embryos (p = 0.0021). No correlation was found between multinucleation and the type of chromosomal abnormality. Limitations, reasons for caution The sample size is the main limitation of the present study. More research with bigger sample size is needed in order to confirm the finding of the present study. Wider implications of the findings: The present study suggests that multinucleated blastomeres during embryo development is not an indication for diminished blastocyst formation and does not affect the clinical outcomes. However, within a sibling embryo population, non-multinucleated embryos tend to be euploid and this finding can be used to advance embryo selection efficiency. Trial registration number Not applicable


2021 ◽  
Vol 116 (3) ◽  
pp. e143
Author(s):  
Yoshihisa Harada ◽  
Emi Fukunaga ◽  
Tomoyo Maeda ◽  
Maki Ikeda ◽  
Reiko Shiba ◽  
...  

2015 ◽  
Vol 27 (1) ◽  
pp. 208
Author(s):  
S. Matoba ◽  
T. Somfai ◽  
T. Nagai ◽  
M. Geshi

Previously, an early first cleavage and a second cleavage after IVF with a normal cleavage pattern defined by even blastomeres without fragments or protrusions was found to be a potent marker for the selection of embryos with high developmental competence (Sugimura et al. 2012 PLoS ONE 7, e36627). The aim of this study was to investigate the effects of bulls and X-sorting of sperm on the ability of these simple noninvasive markers to predict the potency of bovine IVF embryos to develop to the blastocyst stage in vitro. Immature oocytes were matured in TCM199 supplemented with 0.02 armour unit mL–1 FSH and 5% calf serum at 38.5°C in 5% CO2 and 95% air for 22 to 23 h. After maturation, oocytes were inseminated with either of non-sorted frozen-thawed sperm from 3 bulls (A–C) or X-sorted sperm of bull A. Putative zygotes were cultured (IVC) in CR1aa medium supplemented with 5% calf serum and 0.25 mg mL–1 linoleic acid albumin at 38.5°C in 5% CO2, 5% O2, and 90% N2 for 216 h. Embryo kinetics were observed individually by time-lapse cinematography (CCM-1.3Z; Astec, Fukuoka, Japan; Sugimura et al. 2010 Biol. Reprod. 83, 970–978). First and second cleavage kinetics and pattern were categorized according to Sugimura et al. (2012). For each bull, blastocyst development from embryos possessing the following 3 selection markers was compared: (marker 1) the first cleavage within 28 h after IVF, (marker 2) marker 1 combined with 2 even blastomeres without fragments or protrusions, and (marker 3) marker 2 combined with the second cleavage within 50 h after IVF with ≥6 even blastomeres without fragments or protrusions, respectively. Data were analysed by the Yates' corrected chi-square test. A total of 823 oocytes were used in at least 3 replications. When non-sorted sperm was used for IVF, there was not difference (P > 0.05) in total blastocyst formation rates on Day 8 (Day 0 = IVF) among bulls (ranging between 49.5 and 60.8%); however, blastocyst formation rate of embryos generated from X-sorted sperm of bull A (39.5%) was lower (P < 0.05) compared with other groups despite of similar cleavage rates. Embryos having marker 3 criteria developed to the blastocysts stage at significantly higher rates than those having marker 1 criteria in case of non-sorted sperm of bulls A, B, C, and X-sorted sperm of bull A (75.9, 87.0, 90.0, and 75.0% v. 59.5, 62.2, 63.6, and 46.3%, respectively). In groups produced from non-sorted sperm of bulls A, B, C, and X-sorted sperm of bull A, blastocyst development rates of embryos with marker 2 criteria (73.7, 75.0, 90.0, and 65.8%, respectively) were higher (P < 0.05) than those of embryos having marker 1 criteria but did not differ significantly from those with marker 3 criteria. Our results reveal that a first cleavage within 28 h after IVF to 2 even blastomeres without fragments or protrusions are potent predictive markers of the developmental competence of bovine embryos to the blastocyst stage regardless of bulls and sperm sorting.Research was partly supported by JSPS KAKENHI (26450388).


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
N D Munck ◽  
N Nobrega ◽  
A Abdala ◽  
A El-Damen ◽  
A Arnanz ◽  
...  

Abstract Study question Does the limited exposure of embryos outside the incubator, during evaluation and changeover, have an impact on the blastocyst development, blastocyst quality and euploid outcomes? Summary answer Exposure of embryos outside the incubator, negatively impacts the number, quality and euploidy rate of day 5 blastocysts. What is known already The laboratory environment with its culture conditions is one of the crucial elements of the delicate equation to a successful ART outcome. It has been shown that increased fluctuations in the culture conditions have a considerable impact on the number of blastocysts obtained and cycle outcomes. Compared to conventional benchtop incubators, Time Lapse Technology (TLT) incubators capture images of the embryo and allow morphologic and morphokinetic assessment without disturbance during incubation. Several studies have been published comparing the efficiency, safety and outcome performance between conventional and TLT incubators, however, none of them explored the euploid outcomes. Study design, size, duration An observational sibling oocyte study was performed at ART Fertility Clinics, Abu Dhabi between March 2018 and April 2020 and included data of 796 mature oocytes injected from 42 stimulation cycles. Sibling oocytes were randomly split between 2 different incubators: 12 oocytes were assigned to the twelve wells of the EmbryoscopeTM (ES) and the remaining oocytes were cultured in a conventional benchtop incubator, G185 K-System (KS). Participants/materials, setting, methods Embryos from patients with primary or secondary infertility, who underwent ovarian stimulation for ICSI and PGT-A through NGS on trophectoderm biopsies, were eligible. All patients had at least 16 fresh mature oocytes, randomly allocated to two different incubators after ICSI: 503 (63.2%) oocytes were cultured in ES and 293 (36.8%) in KS. The fertilization, cleavage, useable blastocyst and euploid rates, as well as embryo/blastocyst qualities were assessed to evaluate each incubator’s performance. Main results and the role of chance The fertilization and cleavage rates were similar between incubators. Total useable blastocyst rate (64.8% vs 49.6%, p &lt; 0.001) was significantly higher for embryos cultured in ES, mainly due a higher percentage of blastocysts biopsied on day 5 in ES (67.8% vs 57.0%, p = 0.037), with improved quality (p = 0.008). There was no difference in the total euploid rate between ES and KS (59.9% vs 50.4%, p = 0.314), but a significantly higher euploid rate was seen for blastocysts cultured in ES and biopsied on day 5 (63.5% vs 37.4%, p = 0.001). Day 3 embryo quality and total biopsied blastocyst quality was not different between incubators. No difference was observed in the total useable blastocyst development from good (p = 0.0832) and poor (p = 0.112) quality day 3 cleavage stage embryos. However, when stratifying according to the day of blastocyst development, poor quality embryos on day 3 showed superior blastocyst formation on day 5 when cultured in ES (64.1% vs 39.1% for day 5 and 35.9% vs 60.9% for day 6, p = 0.005). Accordingly, blastocyst formation from poor quality embryos on day 3, was shifted to day 6 for embryos cultured in KS. This difference in the day of blastocyst development was not observed for good quality cleavage stage embryos (p = 0.917). Limitations, reasons for caution The current observational study needs confirmation in a prospective trial and should also include the implantation potential of the euploid blastocysts, which was not followed in the current study. A good prognosis population (≥16 mature oocytes) was studied and may not reflect the outcomes in patients with lower oocyte numbers. Wider implications of the findings: This work builds evidence to the solid introduction of the TLT incubators to the clinical routine, as the reduced exposure of embryos outside the incubator – and hence decreased stress - improves the blastocyst development. Trial registration number NA


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