A Benchtop Incubator Prospective Study

ART Fertility Clinics LLC

Status: Enrolling

Conditions: Infertility

Treatments: Other: Culture media preparation and refreshment; Other: Media refreshment Group 1; Other: Insemination and culture; Other: Media refreshment Group 2; Other: Embryo grading

Study type: Observational

Funder types: Other

Identifiers

NCT04969575

2105-ABU-008-IK

Details and patient eligibility

About

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.

Full 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.

Enrollment: 50 estimated patients

Sex: Female

Ages: 18 to 40 years old

Volunteers: Accepts Healthy Volunteers

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 ejaculated sperm >10,000 motile sperm
• Any ovarian stimulation
• Embryos cultured in benchtop incubator

Exclusion criteria

• 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 design

50 participants in 2 patient groups

day 3 refreshment

Description:

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.

Treatment:

Other: Insemination and culture

Other: Media refreshment Group 1

Other: Culture media preparation and refreshment

Other: Embryo grading

day 5 refreshment

Description:

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.

Treatment:

Other: Insemination and culture

Other: Media refreshment Group 2

Other: Culture media preparation and refreshment

Other: Embryo grading