A Time-lapse Monitoring Prospective Study

Exploring Single Step Culture Medium Refreshment Effect on Embryo Ploidy Rate: A Time-lapse Monitoring Prospective Study

While numerous types of commercially available human embryo culture media exist for human blastocyst culture, the impact of culture conditions on blastocyst development and aneuploidy formation is not fully understood. Culture conditions are very important for the success of the in vitro fertilization (IVF) cycle, many of the factors involved in the process have been extensively studied. However, none of the studies investigated the effect on euploid rate in a sibling oocyte design with preimplantation genetic testing for aneuploidy (PGT-A), which requires culture till day 7. In addition, the clinical outcome (implantation) will be investigated in a frozen cycle regimen. Hence, the study will explore which day of media refreshment will result in higher rate of ploidy and would improve clinical outcomes. Investigators aim at exploring the best practice, that would empower the euploid rate through the comparison of refreshing the single-step medium on day 3 or day 5 in a sibling oocyte prospective design.

Study Overview

• Status: Active, not recruiting
• Conditions: Infertility; Aneuploidy
• Intervention / Treatment: Behavioral: media refreshment; Behavioral: embryo grading

Detailed Description

The use of extended embryo culture to the blastocyst stage and PGT-A has enabled identification and selection of embryos with the best developmental potential and improved IVF clinical outcomes. It is considered a routine practice to grade the embryos on day 3 of development, which will require disturbing the culture conditions by annotating the embryos using inverted microscopes. However, the application of Time-lapse technology (TLS) allows annotating the embryos on day 3 without interrupting the culture conditions, especially if TLS is combined with single-step culture medium.

There are several culture factors that can affect embryo development, such as the pH, which is controlled through the concentration of CO2, and also the osmolality, which is the concentration of solute per liter of solution. Evaporation of the solution due to the exposure to high temperature (37°C) can change the concentrations and hence compromise embryo development.

Since single-step medium is designed to sustain embryo development for 5 continuous culture days, while some laboratories culture embryos for 7 days, refreshing the culture medium is inevitable either on day 3 or day 5 of development.

There is no concrete evidence on which of these practices will provide a better environment for embryo development. Moreover, none of the studies investigated the effect of single-step culture medium changeover on day 3 or day 5, on the chromosomal errors (aneuploidy). We aim in this study to explore which day of culture medium refreshment (day 3 or day 5) provides a higher rate of euploid blastocysts.

Summary about data collected. Categorical data will be presented in frequency tables and histograms. Continuous values will be summarized according the mean, standard deviation and confidence interval.

• Study Type: Observational
• Enrollment (Anticipated): 50

Contacts and Locations

• Study Contact: Name: BARBARA LAWRENZ, PhD; Phone Number: 1108 97126528000; Email: shieryl.digma@artfertilityclinics.com
• Study Contact Backup: Name: SHIERYL DIGMA, RN; Phone Number: 1061 97126528000; Email: shieryl.digma@artfertilityclinics.com

Study Locations

• United Arab Emirates
  • Abu Dhabi, United Arab Emirates
    • ART Fertility Clinics
  • Abu Dhabi, United Arab Emirates
    • Ibrahim Elkhatib

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 40 years (Adult)
• Accepts Healthy Volunteers: N/A
• Genders Eligible for Study: Female

• Sampling Method: Non-Probability Sample

Study Population

Patients undergoing a fertility treatment due to primary or secondary infertility at ART Fertility Clinics.

Description

Inclusion Criteria:

• Patients with a minimum of 8 - 10 follicles during ovarian stimulation
• Patients with a minimum of 8 mature oocytes
• PGT-A
• ICSI
• BMI<35
• Age 18- 40
• Fresh or frozen ejaculatory sperm >10,000 motile sperm
• Any Ovarian stimulation
• Embryos cultured in Time-lapse system

Exclusion Criteria:

• PGT-M or PGT-SR
• Previous history of embryo arrest
• Endometriosis
• Hydrosalpinx
• History of uterine abnormality or previous uterine surgery
• History of previous treatment which may impact the ovarian reserve (adnexal surgery, chemotherapy, radiation…)

Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
day 3; embryo grading and culture refreshment will be done 67-69 hours post insemination	Behavioral: media refreshment; 20µL of culture medium is aspirated using Eppendorf PCR tips, and freshly, overnight equilibrated 20µL global total LP will be added to the same well; Behavioral: embryo grading; embryo grading should be done prior to media refreshment
day 5; embryo grading and culture medium refreshment will be done 114-118 hours post insemination	Behavioral: media refreshment; 20µL of culture medium is aspirated using Eppendorf PCR tips, and freshly, overnight equilibrated 20µL global total LP will be added to the same well; Behavioral: embryo grading; embryo grading should be done prior to media refreshment

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
blastocyst ploidy	determined after biopsy of trophectoderm cells, taken from the blastocyst on day 5, 6 or 7 from development.	3 days

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
cycle ploidy rate	the number of euploid embryos in the group divided by the total number of 2PN	1 day
mosaicism frequency	defined as the rate of mosaic errors in aneuploid embryos. This includes full mosaic embryos or aneuploid embryos with 1 or more mosaic chromosome aberration	1 day
blastocyst quality at time of biopsy	Four Embryo Quality Scores (EQ1, 2, 3, 4) based on the expansion stage of the blastocyst, the grade of the inner cell mass and the grade of the trophectoderm	1 day
blastulation rate	ability of the embryo to form a cavity (=to blastulate) 118hours post ICSI. Number of blastocysts divided by the number of 2PN	5 days

Collaborators and Investigators

• Sponsor: ART Fertility Clinics LLC
• Investigators: Principal Investigator: IBRAHIM ELKHATIB, ART Fertility Clinics LLC

Publications and helpful links

General Publications

• Bavister BD, Poole KA. Duration and temperature of culture medium equilibration affect frequency of blastocyst development. Reprod Biomed Online. 2005 Jan;10(1):124-9. doi: 10.1016/s1472-6483(10)60813-9.
• Costa-Borges N, Belles M, Meseguer M, Galliano D, Ballesteros A, Calderon G. Blastocyst development in single medium with or without renewal on day 3: a prospective cohort study on sibling donor oocytes in a time-lapse incubator. Fertil Steril. 2016 Mar;105(3):707-713. doi: 10.1016/j.fertnstert.2015.11.038. Epub 2015 Dec 12.
• Krasnopolskaya KV, Beketova AN, Sesina NI, capital ES, Cyrillichinchenko NK, Badalyan GV, Sudarikova NM, Bocharova TV, Zakharchenko EO. The effect of short-term disturbance of day 3 embryo culture on the development and implantation. Gynecol Endocrinol. 2019;35(sup1):1-4. doi: 10.1080/09513590.2019.1632083.

Helpful Links

• Water evaporation and osmolarity change of human embryo culture media in humid or in dry culture systems

Study record dates

Study Major Dates

• Study Start (Actual): July 6, 2021
• Primary Completion (Anticipated): June 30, 2023
• Study Completion (Anticipated): December 30, 2023

Study Registration Dates

• First Submitted: May 20, 2021
• First Submitted That Met QC Criteria: May 20, 2021
• First Posted (Actual): May 25, 2021

Study Record Updates

• Last Update Posted (Actual): March 24, 2023
• Last Update Submitted That Met QC Criteria: March 23, 2023
• Last Verified: March 1, 2023

More Information

Terms related to this study

• Keywords: infertility; aneuploidy; embryo; culture media; ivf; embryoscope
• Additional Relevant MeSH Terms: Pathologic Processes; Chromosome Aberrations; Infertility; Aneuploidy
• Other Study ID Numbers: 2103-ABU-003-IK

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No