Is the AMH Intrafollicular Level a Predictor of the Ploidy Status of the Blastocyst?

Study Overview
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Brief Summary
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To investigate, whether on the day of egg retrieval, after ovarian stimulation for ICSI (intracytoplasmic sperm injection), there is a correlation between the intrafollicular AMH (Anti-Müllerian hormone) levels and the ploidy status of the blastocyst.

Detailed Description
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In clinical practice, AMH is used as a diagnostic and/or prognostic marker in women in association with ovulation induction and in various pathophysiological conditions. This study looks to investigate if the hormonal environment that surrounds an oocyte has an impact on the ploidy status of the blastocyst. Nowadays, using NGS (Next Generation Sequencing) platform for analysis of 23 pairs of chromosomes for Preimplantation Genetic Testing on blastocysts, the ploidy status of the embryo is more adequately assessed. Furthermore, embryo culture to blastocyst will provide further information about embryo quality and possibilities of implantation. To have complete information between the serum hormones, follicular hormones, embryo development and ploidy will provide to clinicians, embryologist and patients further information on embryo selection and adequate ovarian stimulation protocols. The database will be exported from the clinical information manager, VRepro, to a table in excel format through a database-based query system. The exported data will be duly codified to protect the clinical and personal information of the participants. Prior to the statistical study, an exploratory data analysis will be carried out to review the quality of the information extracted.

Official Title
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Is the AMH Intrafollicular Level a Predictor of the Ploidy Status of the Blastocyst? A Prospective Study

Conditions
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Infertility, Female, Infertility

Intervention / Treatment
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* Procedure: Oocyte Pick Up - Out of Study
* Procedure: Denudation - Out of Study
* Procedure: ICSI
* Procedure: Culture
* Genetic: Biopsy
* Procedure: Vitrification
* Procedure: OPU - Study Group
* Procedure: Denudation - Study Group


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: At least one follicle from each ovary at day of trigger Age from 18 to 40 years old Normal menstrual cycle of 25-35 days Poor, normal and high response will be included First follicle punctured from each ovary will be consider for inclusion: if the COC (Cumulus Oocyte Complex) is present in clear FF (Follicular fluid), the FF will be collected separately for further analysis and the oocyte included in the study Exclusion Criteria: Very hematic follicular fluid If the COC is not found in the follicular fluid Azoospermia Known chromosomal abnormalities Severe OAT (Oligoasthenoteratozoospermia)

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 40 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Study Group<br> | Procedure: ICSI<br>* One oocyte injected at a time.<br>Procedure: Culture<br>* One embryo per culture drop. Individual follow-up of the embryo development.<br>Genetic: Biopsy<br>* One embryo per culture drop.<br>Procedure: Vitrification<br>* One embryo per vitrification straw.<br>Procedure: OPU - Study Group<br>* Measure the size of the follicle and aspirate only one follicle and flush. The needle before moving to the second ovary. Oocyte included in the study will be cultured separately (each ovary separately too).<br>Procedure: Denudation - Study Group<br>* Oocyte included in the study will be denuded separately. Maturational stages are recorded per follicle.<br>|
| Out of Study Group<br> | Procedure: Oocyte Pick Up - Out of Study<br>* Aspiration of follicles. search for the oocyte(s) All oocytes are grouped in one dish.<br>Procedure: Denudation - Out of Study<br>* All oocytes are denuded together. Maturational stages are recorded.<br>Procedure: ICSI<br>* One oocyte injected at a time.<br>Procedure: Culture<br>* One embryo per culture drop. Individual follow-up of the embryo development.<br>Genetic: Biopsy<br>* One embryo per culture drop.<br>Procedure: Vitrification<br>* One embryo per vitrification straw.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Oocyte maturity | Calculated as percentage transformation of an oocyte from prophase I to metaphase II | 1 day | 
| Fertilization | Percentage of mature oocytes - metaphase II (MII) that become 2 pronucleii embryos (2PN) | 1 day | 
| Blastulation | Ability of the embryo to form a cavity (=to blastulate) 114-118 hours post ICSI. Number of blastocysts divided by the number of 2PN | 1 day | 
| Blastocyst quality | 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. | 6 days | 
| Ploidy status | Determined after biopsy of trophectoderm cells, taken from the blastocyst on day 5, 6 or 7 from development | 14 days | 
| Mitochondria status | Measured as mtDNA ratio, the ratio of mitochondrial DNA (mtDNA) read counts to autosomal (chromosome 1-22) DNA read counts | 14 days | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| AMH in the follicular fluid. | To evaluate if there is an association between the levels of serum AMH on day 2-3 of the stimulation, AMH day of OPU and the intra-follicular AMH. | 1 day | 
| FSH in the follicular fluid. | To evaluate if there is an association between serum FSH on day 2-3 of the stimulation, serum FSH day of OPU and the intra-follicular FSH. | 1 day | 
| LH in the follicular fluid. | To evaluate if there is an association between serum LH on day 2-3 of the stimulation, serum LH day of OPU and the intra-follicular LH. | 1 day | 
| E2 in the follicular fluid. | To evaluate if there is an association between serum E2 on day 2-3 of the stimulation, serum E2 day of OPU and the intra-follicular E2. | 1 day | 
| P4 in the follicular fluid. | To evaluate if there is an association between serum P4 on day 2-3 of the stimulation, serum P4 day of OPU and the intra-follicular P4. | 1 day | 
| Inhibin B in the follicular fluid. | To evaluate if there is an association between serum Inhibin B on day 2-3 of the stimulation, serum Inhibin B day of OPU and the intra-follicular Inhibin B. | 1 day | 
| AMH in serum day of OPU | To evaluate if there is an association between the levels of serum AMH on day 2-3 of the stimulation, AMH day of OPU and the intra-follicular AMH. | 1 day | 
| FSH in serum day of OPU | To evaluate if there is an association between serum FSH on day 2-3 of the stimulation, serum FSH day of OPU and the intra-follicular FSH. | 1 day | 
| LH in serum day of OPU | To evaluate if there is an association between serum LH on day 2-3 of the stimulation, serum LH day of OPU and the intra-follicular LH. | 1 day | 
| E2 in serum day of OPU | To evaluate if there is an association between serum E2 on day 2-3 of the stimulation, serum E2 day of OPU and the intra-follicular E2. | 1 day | 
| P4 in serum day of OPU | To evaluate if there is an association between serum P4 on day 2-3 of the stimulation, serum P4 day of OPU and the intra-follicular P4. | 1 day | 
| Inhibin B in serum day of OPU | To evaluate if there is an association between serum Inhibin B on day 2-3 of the stimulation, serum Inhibin B day of OPU and the intra-follicular Inhibin B. | 1 day | 
| AMH in serum day of day 2/3 of stimulation. | To evaluate if there is an association between the levels of serum AMH on day 2-3 of the stimulation, AMH day of OPU and the intra-follicular AMH. | 1 day | 
| FSH in serum day of day 2/3 of stimulation. | To evaluate if there is an association between serum FSH on day 2-3 of the stimulation, serum FSH day of OPU and the intra-follicular FSH. | 1 day | 
| LH in serum day of day 2/3 of stimulation. | To evaluate if there is an association between serum LH on day 2-3 of the stimulation, serum LH day of OPU and the intra-follicular LH. | 1 day | 
| E2 in serum day of day 2/3 of stimulation. | To evaluate if there is an association between serum E2 on day 2-3 of the stimulation, serum E2 day of OPU and the intra-follicular E2. | 1 day | 
| P4 in serum day of day 2/3 of stimulation. | To evaluate if there is an association between serum P4 on day 2-3 of the stimulation, serum P4 day of OPU and the intra-follicular P4. | 1 day | 
| Inhibin B in serum day of day 2/3 of stimulation. | To evaluate if there is an association between serum Inhibin B on day 2-3 of the stimulation, serum Inhibin B day of OPU and the intra-follicular Inhibin B. | 1 day | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Estradiol, FSH, LH, AMH, Ploidy, Chromosomes, Infertility, Inhibin B, Progesterone