InfertilityNCT04969575
A Benchtop Incubator Prospective Study
An observational study of Infertility, run by ART Fertility Clinics LLC at 1 site in United Arab Emirates.
Trial summary
The use of extended embryo culture to the blastocyst stage and preimplantation genetic testing for aneuploidy (PGT-A) has enabled identification and selection of embryos with the best developmental potential and improved in vitro fertilization (IVF) clinical outcomes. Read more about this study
- Who is it for?
- Ages 18 to 40 years, women onlyHealthy volunteers can take part
- Conditions the study is for
- Infertility
- What is being tested?
- Insemination and culture, Media refreshment Group 1 and Culture media preparation and refreshment
- How many people
- 50 participants
- Where
- 1 siteUnited Arab Emirates
- Study dates
- 15 Jul 2021to 31 Dec 2023
- Phase
- Observational
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 40 years
- Women only
- Healthy volunteers accepted
- Adult
Study population: The study is eligible to all patients, meeting the inclusion criteria, undergoing a fertility treatment with PGT-A analysis at the blastocyst stage
Who may be able to join
- 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
Show all 9Show fewer
- Fresh or frozen ejaculated sperm >10,000 motile sperm
- Any ovarian stimulation
- Embryos cultured in benchtop incubator
Who cannot join
- PGT-M or PGT-SR
- Previous history of embryo arrest
- Endometriosis
- History of previous treatment which may impact the ovarian reserve (adnexal surgery, chemotherapy, radiation…)
Trial locations and contacts
1 site in 1 country.
United Arab Emirates1 site, 1 recruiting
- Recruiting
ART Fertility Clinics LLC
Abu Dhabi, 60202
Barbara Lawrenz, PhD, +97102 652 8000 ext. 1108, shieryl.digma@artfertilityclinics.com
Shieryl Digma, RN, +97102 652 8000 ext. 1122, shieryl.digma@artfertilityclinics.com
About this study
Official title: Exploring Single Step Culture Medium Refreshment Effect on Embryo Ploidy Rate: A Benchtop Incubator Prospective Study
The use of extended embryo culture to the blastocyst stage and preimplantation genetic testing for aneuploidy (PGT-A) has enabled identification and selection of embryos with the best developmental potential and improved in vitro fertilization (IVF) clinical outcomes. While numerous types of commercially available human embryo culture media exist for culture to the blastocyst stage, the impact of culture conditions on blastocyst development and aneuploidy formation is not well understood.
Culture conditions are very important for the success of the IVF cycle, many of the factors involved in the process have been extensively studied, including the use of sequential culture medium vs single step culture medium, humidified incubators vs dry incubators and refreshing the medium on day 3 vs uninterrupted culture till day 5. However, none of the studies investigated the effect on euploid rate in a sibling oocyte design with PGT-A, which requires culture till day 7.
Read the detailed description
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. Extended culture to day 7 is required in order to perform trophectoderm biopsy required for PGT-A. 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.
To reduce the rate of evaporation, culture droplets are covered with a layer of mineral oil. Humidified incubators can also reduce the rate of evaporation. However, in the case of dry incubators, such as benchtop incubators, replenishing the culture medium at least once -by moving the embryos to a new culture dish- during these 7 days of culture might be the solution.
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/mosaicism). Investigators aim in this prospective sibling oocyte study, to explore which day of culture medium refreshment (day 3 or day 5) provides a higher rate of euploid blastocysts.
- Conditions
- Infertility
- Being tested
- Insemination and culture, Media refreshment Group 1, Culture media preparation and refreshment, Embryo grading, Media refreshment Group 2
- Keywords
- IVF, incubator, aneuploidy, embryoscope, embryo, ICSI
- Medical subject headings
- Infertility, Aneuploidy
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Insemination and cultureOther
All MII oocytes will be inseminated by ICSI 39-41 hours post final oocyte maturation trigger.
Media refreshment Group 1Other
Zygote scoring on day 1 (18-20hpi). Culture medium refreshment will be done 67-69 hours post insemination (Day3).
Culture media preparation and refreshmentOther
30µL of culture medium (Global total LP is added to each well of the 8 wells of LifeGlobal GPS Dish (EGPS-010) and 50µL to the center 3 drops.
Dishes will be equilibrated overnight in a CO2 incubator.
Embryo gradingOther
Should be done prior to media refreshment. For group 2, embryo grading and media refreshment is done prior to blastocyst biopsy.
Media refreshment Group 2Other
Zygote scoring on day 1 (18-20hpi). Culture media refreshment will be done 114-118 hours post insemination (Day5).
| Study group | Role | What they receive |
|---|---|---|
| day 3 refreshment | All MII oocytes will be inseminated by ICSI 39-41 hours post final oocyte maturation trigger.
Following ICSI, oocytes will be cultured in 2-4 culture dishes placed in 2 separate chambers.
Media refreshment: Zygote scoring on day 1 (18-20hpi). Culture medium refreshment will be done 67-69 hours post insemination (Day3). Embryo grading will not be done day2-5. Culture media preparation and refreshment: 30µL of culture medium (Global total LP is added to each well of the 8 wells of LifeGlobal GPS Dish (EGPS-010) and 50µL to the center 3 drops.
Dishes will be equilibrated overnight in a CO2 incubator. Embryo grading should be done prior to media refreshment. Time out of the incubator during media refreshment will be assessed on day 3 and day 5.
One operator should perform the ICSI. In case of multiple operators, each operator's injected oocytes will be split between the 2 groups. Receives: Other: Insemination and culture; Other: Media refreshment Group 1; Other: Culture media preparation and refreshment; Other: Embryo grading | |
| day 5 refreshment | All MII oocytes will be inseminated by ICSI 39-41 hours post final oocyte maturation trigger.
Following ICSI, oocytes will be cultured in 2-4 culture dishes placed in 2 separate chambers.
Media refreshment and embryo grading timings: Zygote scoring on day 1 (18-20hpi). Embryo grading and culture media refreshment will be done 114-118 hours post insemination (Day5).
Culture media preparation and refreshment: 30µL of culture medium (Global total LP is added to each well of the 8 wells of LifeGlobal GPS Dish (EGPS-010) and 50µL to the center 3 drops. Dishes will be equilibrated overnight in a CO2 incubator.
Embryo grading and media refreshment is done prior to blastocyst biopsy. Time out of the incubator during media refreshment will be assessed on day 3 and day 5.
One operator should perform the ICSI. In case of multiple operators, each operator's injected oocytes will be split between the 2 groups. Receives: Other: Insemination and culture; Other: Culture media preparation and refreshment; Other: Embryo grading; Other: Media refreshment Group 2 |
What is being measured
The primary outcome is the main question the study is designed to answer.
Blastocyst ploidy
determined after biopsy of trophectoderm cells, taken from the blastocyst on day 5, 6 or 7 from development
Measured: 3 days
Secondary outcomes (4)
Cycle ploidy rate
the number of euploid embryos in the group divided by the total number of 2PN (both groups)
Measured: 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
Measured: 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.
Measured: 1 day
Blastulation rate
ability of the embryo to form a cavity (=to blastulate) 114-118 hours post ICSI. Number of blastocysts divided by the number of 2PN
Measured: 5 days
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted20 Jul 2021
- Study start15 Jul 2021
- Primary completion30 Jun 2023 (estimated)
- Study completion31 Dec 2023 (estimated)
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (5)Papers the sponsor has linked to this study.
- Other trials for InfertilityStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Observational
- Observational model
- Cohort
- Time perspective
- Retrospective
- Enrolment
- 50 participants (estimated)
Sponsor and investigators
- Lead sponsor
- ART Fertility Clinics LLC
- Sponsor type
- University, hospital or other
- Responsible party
- Principal investigator, Ibrahim Elkhatib, ART Fertility Clinics LLC
- Investigators
- IBRAHIM ELKHATIB, Principal investigator, ART Fertility Clinics LLC
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT04969575
- Sponsor's protocol ID
- 2105-ABU-008-IK
- Status last verified
- Mar 2023
- First submitted
- 11 Jul 2021
- Last updated
- 24 Mar 2023
- Results posted
- No
- Data monitoring committee
- Yes
- FDA-regulated drug
- No
- FDA-regulated device
- No
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.
PubMed 15705308Costa-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.
PubMed 26690010Pena F, Davalos R, Rechkemmer A, Ascenzo A, Gonzales M. Embryo development until blastocyst stage with and without renewal of single medium on day 3. JBRA Assist Reprod. 2018 Mar 1;22(1):49-51. doi: 10.5935/1518-0557.20180012.
PubMed 29318868Krasnopolskaya 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.
PubMed 31532309Elkhatib I, Bayram A, Abdala A, Hourani N, Ferracuti V, Siqueira S, Kalafat E, Fatemi HM, Nogueira D. Day 3 versus Day 5 refreshment of single-step medium for embryos cultured in a benchtop dry incubator: a randomized sibling oocyte study. Reprod Biomed Online. 2026 Mar;52(3):105288. doi: 10.1016/j.rbmo.2025.105288. Epub 2025 Oct 9.
PubMed 41702273
Study at a glance
From the registry record, last updated by the sponsor on 24 Mar 2023. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Exploring Single Step Culture Medium Refreshment Effect on Embryo Ploidy Rate: A Benchtop Incubator Prospective Study
- ClinicalTrials.gov ID
- NCT04969575
- Acronym / sponsor ID
- 2105-ABU-008-IK
- Conditions
- Infertility
- Keywords
- IVF, incubator, aneuploidy, embryoscope, embryo, ICSI
- Study type
- Observational
- Recruitment status
- Unknown status
- Enrolment
- 50 participants (estimated)
- Lead sponsor
- ART Fertility Clinics LLCUniversity, hospital or other
- Study start
- 15 Jul 2021 (actual)
- Primary completion
- 30 Jun 2023 (estimated)
- Study completion
- 31 Dec 2023 (estimated)
- First posted
- 20 Jul 2021 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
The use of extended embryo culture to the blastocyst stage and preimplantation genetic testing for aneuploidy (PGT-A) has enabled identification and selection of embryos with the best developmental potential and improved in vitro fertilization (IVF) clinical outcomes. While numerous types of commercially available human embryo culture media exist for culture to the blastocyst stage, the impact of culture conditions on blastocyst development and aneuploidy formation is not well understood.
Culture conditions are very important for the success of the IVF cycle, many of the factors involved in the process have been extensively studied, including the use of sequential culture medium vs single step culture medium, humidified incubators vs dry incubators and refreshing the medium on day 3 vs uninterrupted culture till day 5. However, none of the studies investigated the effect on euploid rate in a sibling oocyte design with PGT-A, which requires culture till day 7.
Detailed description
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. Extended culture to day 7 is required in order to perform trophectoderm biopsy required for PGT-A. 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.
To reduce the rate of evaporation, culture droplets are covered with a layer of mineral oil. Humidified incubators can also reduce the rate of evaporation. However, in the case of dry incubators, such as benchtop incubators, replenishing the culture medium at least once -by moving the embryos to a new culture dish- during these 7 days of culture might be the solution.
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/mosaicism). Investigators aim in this prospective sibling oocyte study, to explore which day of culture medium refreshment (day 3 or day 5) provides a higher rate of euploid blastocysts.
Study design
- Observational model
- Cohort
- Time perspective
- Retrospective
- Number of arms
- 2
| Arm | Type | Receives |
|---|---|---|
| day 3 refreshment | – | Other: Insemination and culture, Other: Media refreshment Group 1, Other: Culture media preparation and refreshment, Other: Embryo gradingAll MII oocytes will be inseminated by ICSI 39-41 hours post final oocyte maturation trigger. Following ICSI, oocytes will be cultured in 2-4 culture dishes placed in 2 separate chambers. Media refreshment: Zygote scoring on day 1 (18-20hpi). Culture medium refreshment will be done 67-69 hours post insemination (Day3). Embryo grading will not be done day2-5. Culture media preparation and refreshment: 30µL of culture medium (Global total LP is added to each well of the 8 wells of LifeGlobal GPS Dish (EGPS-010) and 50µL to the center 3 drops. Dishes will be equilibrated overnight in a CO2 incubator. Embryo grading should be done prior to media refreshment. Time out of the incubator during media refreshment will be assessed on day 3 and day 5. One operator should perform the ICSI. In case of multiple operators, each operator's injected oocytes will be split between the 2 groups. |
| day 5 refreshment | – | Other: Insemination and culture, Other: Culture media preparation and refreshment, Other: Embryo grading, Other: Media refreshment Group 2All MII oocytes will be inseminated by ICSI 39-41 hours post final oocyte maturation trigger. Following ICSI, oocytes will be cultured in 2-4 culture dishes placed in 2 separate chambers. Media refreshment and embryo grading timings: Zygote scoring on day 1 (18-20hpi). Embryo grading and culture media refreshment will be done 114-118 hours post insemination (Day5). Culture media preparation and refreshment: 30µL of culture medium (Global total LP is added to each well of the 8 wells of LifeGlobal GPS Dish (EGPS-010) and 50µL to the center 3 drops. Dishes will be equilibrated overnight in a CO2 incubator. Embryo grading and media refreshment is done prior to blastocyst biopsy. Time out of the incubator during media refreshment will be assessed on day 3 and day 5. One operator should perform the ICSI. In case of multiple operators, each operator's injected oocytes will be split between the 2 groups. |
Who may qualify
- Ages
- 18 to 40 years
- Sex
- Female only
- Healthy volunteers
- Accepted
- Age groups
- Adult
- Study population
- The study is eligible to all patients, meeting the inclusion criteria, undergoing a fertility treatment with PGT-A analysis at the blastocyst stage
Key inclusion criteria (9 listed)
- Patients with a minimum of 8 - 10 follicles during ovarian stimulation
- Patients with a minimum of 8 mature oocytes
- PGT-A
- ICSI
- BMI<35
Key exclusion criteria (4 listed)
- PGT-M or PGT-SR
- Previous history of embryo arrest
- Endometriosis
- History of previous treatment which may impact the ovarian reserve (adnexal surgery, chemotherapy, radiation…)
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Insemination and cultureAll MII oocytes will be inseminated by ICSI 39-41 hours post final oocyte maturation trigger. | Other | day 3 refreshment, day 5 refreshment |
| Media refreshment Group 1Zygote scoring on day 1 (18-20hpi). Culture medium refreshment will be done 67-69 hours post insemination (Day3). | Other | day 3 refreshment |
| Culture media preparation and refreshment30µL of culture medium (Global total LP is added to each well of the 8 wells of LifeGlobal GPS Dish (EGPS-010) and 50µL to the center 3 drops. Dishes will be equilibrated overnight in a CO2 incubator. | Other | day 3 refreshment, day 5 refreshment |
| Embryo gradingShould be done prior to media refreshment. For group 2, embryo grading and media refreshment is done prior to blastocyst biopsy. | Other | day 3 refreshment, day 5 refreshment |
| Media refreshment Group 2Zygote scoring on day 1 (18-20hpi). Culture media refreshment will be done 114-118 hours post insemination (Day5). | Other | day 5 refreshment |
Primary outcome
Blastocyst ploidy
Time frame: 3 days
determined after biopsy of trophectoderm cells, taken from the blastocyst on day 5, 6 or 7 from development
4 secondary outcomes
Cycle ploidy rate
Time frame: 1 day
Mosaicism Frequency
Time frame: 1 day
Blastocyst quality at time of biopsy
Time frame: 1 day
Blastulation rate
Time frame: 5 days
Results and safety
Results are not available in the source record.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 24 Mar 2023.
- Background: PubMed 15705308Bavister 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.
- Background: PubMed 26690010Costa-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.
- Background: PubMed 29318868Pena F, Davalos R, Rechkemmer A, Ascenzo A, Gonzales M. Embryo development until blastocyst stage with and without renewal of single medium on day 3. JBRA Assist Reprod. 2018 Mar 1;22(1):49-51. doi: 10.5935/1518-0557.20180012.
- Background: PubMed 31532309Krasnopolskaya 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.
- Derived from this study: PubMed 41702273Elkhatib I, Bayram A, Abdala A, Hourani N, Ferracuti V, Siqueira S, Kalafat E, Fatemi HM, Nogueira D. Day 3 versus Day 5 refreshment of single-step medium for embryos cultured in a benchtop dry incubator: a randomized sibling oocyte study. Reprod Biomed Online. 2026 Mar;52(3):105288. doi: 10.1016/j.rbmo.2025.105288. Epub 2025 Oct 9.
- 1\. Mori, C., Kuwayama, M., Silber, S., Kagawa, N., Kato, H. Water evaporation and osmolality change of human embryo culture media in humid or in dry culture systems. Fertil. Steril. 2010; 94: s151Link provided by the sponsor.
Related trials for Infertility
RecruitingObservational study
Safety and Efficacy of Corifollitropin Alfa N02 Injection in Elderly Women Undergoing Assisted Reproductive Technology (ART)
Infertility
Active, not recruiting
Automation of the In Vitro Fertilization Laboratory
Infertility, ART
RecruitingObservational study
Inherited Reproductive Disorders
Genetic Disorder, Infertility
RecruitingObservational study
Biorepository in Participants Who Undergo OTC for Gonadotoxic Therapy
Acute Ovarian Failure, Infertility
Not yet recruiting
Implementing IVM for Fertility Treatment in Clinical Practice
Infertility
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
IBRAHIM ELKHATIB principal investigator
ART Fertility Clinics LLC
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Infertility, run by ART Fertility Clinics LLC at 1 site in United Arab Emirates.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 9 inclusion and 4 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: status not confirmed, as of the sponsor's last update on 24 Mar 2023. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in United Arab Emirates. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
Results are not available in the source record.
How can a doctor refer a patient or contact the research team?
Contact BARBARA LAWRENZ, PhD (97126528000 ext. 1108, barbara.lawrenz@artfertilityclinics.com), or a site near the patient. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Blastocyst ploidy (time frame: 3 days). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT04969575, which also lists 5 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 24 Mar 2023.