Cumulus Cell Mitochondrial Activity as a Non-invasive Marker of Embryo Quality (FLIM)

Study Overview
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Brief Summary
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This study aims to test the extent of association of FLIM parameters of cumulus cells with clinical outcomes (clinical pregnancy, ongoing pregnancy at 12 weeks, and live birth) of the embryos that result from the enclosed oocytes.

Detailed Description
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This study aims to test the extent of association of FLIM parameters of cumulus cells with clinical outcomes (clinical pregnancy, ongoing pregnancy at 12 weeks, and live birth) of the embryos that result from the enclosed oocytes. Participants will be consented to and pre-signed following standard regimens of IVFMD, My Duc Hospital and IVFMD PN, My Duc Phu Nhuan Hospital. The standard is for patients with at least 8 follicles ≥ 10 mm on ultrasonography at the induced ovulation date. Cumulus samples (CCs) will be collected following the protocol on the day of oocyte pick up (OPU), being assigned with the same number as the oocyte from which it was collected, and vitrified with a Cryotech kit until FLIM measurements are performed. Insemination will be performed by using ICSI, 3 - 4 hours after oocyte retrieval. A sample of CCs will be sliced off the cumulus mass of each oocyte judged to be mature based on the cumulus mass disposition (absence of tight corona layers). The remaining cumulus cells will be stripped from the OCCs using hyaluronidase. Only mature oocytes will be inseminated. The fertilization check will be performed under an inverted microscope for a period of 16-18 hours after insemination. On day 3, embryo evaluation will be performed at a fixed time point 66±2 hours after fertilization and, for day 5/6 transfers at 116±2 hours or 140±2 hours, respectively, using the Istanbul consensus. Freeze-all strategy for day 5 embryo will be applied and the first frozen embryo transfer cycle is single embryo transfer. On the other hand, the investigators will stratify embryos into two groups based on the measured FLIM parameters from their associated cumulus cells, corresponding to the high 35% and low 65% of metabolic scores. The investigators aim for embryos in the high metabolic scoring group to have a 10% higher live birth rate than controls. Assuming a current live birth rate of 32%, this corresponds to a hoped-for 42% live birth rate in the high group (and a 25% live birth rate in the low group). A total of 450 patients are required to detect an effect of this size (power 0.90, two-sided alpha 5%). The primary endpoint of the study will be the live birth rate after the first embryo transfer of the started treatment cycle.

Official Title
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Cumulus Cell Mitochondrial Activity as a Non-invasive Marker of Embryo Quality

Conditions
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IVF/ICSI, Frozen Embryo Transfer, Cumulus Cell, Fluorescence Lifetime Imaging Microscopy

Intervention / Treatment
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* Genetic: FLIM parameters


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Having any number of IVF/ICSI attempts Ovarian stimulation with any stimulation protocol Patients with at least 8 follicles ≥ 10 mm on ultrasonography at induced ovulation date Agreed to freeze-all day 5-6 embryos Agreed to have a single day 5-6 embryo transfer. Agreed to participate in the research (signed the consent form) Exclusion Criteria: In vitro maturation (IVM) cycles Cycles using testicular biopsy or epididymal sperm Cycles not having an oocyte freeze-all cycle Cycles using standard insemination Participating in another research

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Infertile women have IVF/ICSI cycle<br>All Vietnamese infertile women who have IVF/ICSI cycle with at least 8 follicles ≥ 10 mm on ultrasonography at induced ovulation date agreed to freeze-all day 5-6 embryos at IVFMD and IVFMD PN will be enrolled in the study. | Genetic: FLIM parameters<br>* Cumulus samples (CCs) will be collected following the protocol on the day of oocyte pick up (OPU). CCs are frozen by the cryo-tech method, then stored in liquid nitrogen at -196oC. Information of CC samples and processed date/time is recorded. After all, samples are collected, they will be transferred to The Needleman Lab, Northwest Lab Building 359,52 Oxford Street Cambridge MA 02138 to measure mitochondria metabolism using FLIM.<br>* Other names: fluorescence lifetime imaging microscopy;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Live birth rate after one frozen embryo transfer cycle | The birth of at least one newborn, after 24 week's gestation that exhibits any sign of life, such as respiration, heartbeat, umbilical pulsation, or movement of voluntary muscles | After 24 week's gestation | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Positive pregnancy test | Positive pregnancy test is defined as a serum human chorionic gonadotropin level greater than 25 mIU/mL after2 weeks after embryo placement of the first transfer | At 2 weeks after embryo placement | 
| Clinical pregnancy | The presence of at least one gestational sac on ultrasound at 7 week's gestation with the detection of heartbeat activity, after the completion of the first transfer | 7 week's gestation | 
| Implantation rate | Implantation rate is defined as the number of gestational sacs per number of embryos transferred 3 weeks after the first transfer | 3 weeks after the first transfer | 
| Ongoing pregnancy | Having at least 1 gestational sac on ultrasound at 12 weeks' gestation with heart beat activity | At 10 weeks after embryo placement | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Cumulus cell, Day 5/6 frozen embryo, Fluorescence lifetime imaging microscopy, FLIM, IVF/ICSI