Co-incubation of Oocytes With Sperm: Defining the Optimal Incubation Time

Study Overview
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Brief Summary
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The current pilot study aims to evaluate the fertilization rates between sibling oocytes subjected to short incubation (2h) versus overnight incubation (16-20h). As secondary objectives, the abnormal fertilization, embryo development, blastocyst biopsy and euploid rates will be analyzed.

Detailed Description
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All patients for whom at least 6 cumulus oocytes complexes (COCs) are assigned to IVF, with normal sperm parameters, are eligible for the study. An electronically generated randomization list will allocate the first half of the oocytes to one arm and the other half of the oocytes to the second arm. In case an odd number of oocytes will be inseminated, one extra oocyte will be allocated to the first arm. In this prospective observational pilot study, the fertilization potential of 40 patients will be evaluated.

Official Title
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Co-incubation of Oocytes With Sperm: Defining the Optimal Incubation Time

Conditions
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Infertility

Intervention / Treatment
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* Other: 2h exposure to sperm for IVF
* Other: overnight exposure to sperm for IVF


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Sperm parameters Sperm concentration before capacitation: >15 million per ml (WHO) Total motility (PR+NP %): >40 (WHO) Progressive motility (PR %):>32 (WHO) Sperm concentration after capacitation: >0.6 million per ml (not WHO defined) Progressive motility (PR %):>65 (WHO) ≥6 COCs assigned to IVF BMI ≤35 kg/m2 Female age 18 to ≤ 43 years All ovarian stimulation protocols Fresh ejaculates Abstinence duration 2-5 days Presence or absence of sperm morphology data: as we do not have a diagnostic sperm analysis for all patients, the presence or absence of >4% normal morphology (WHO) will not be taken into account, even with known low (<4%) normal morphology Couples requesting Preimplantation Genetic Testing for Aneuploidies Arab population Exclusion Criteria: If the volume to be added after IVF is insufficient to perform IVF on all needed oocytes Presence of >1 million per ml round cells in the ejaculate If a couple's previous cycle was included in the study Previous history of fertilization failure Globozoospermia PCO according to the Rotterdam criteria Progesterone >1.5 ng/ml on the day of trigger

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 43 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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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Oocytes of one patient will be evenly and randomly distributed between two timings of exposure to sperm: 2h exposure versus overnight exposure (16-20h)
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: IVF: 2h exposure<br>Half of a patients' oocytes will be subjected to 2h exposure to sperm | Other: 2h exposure to sperm for IVF<br>* Oocytes will only be briefly (2h) exposed to progressive motile sperm for insemination<br>* Other names: short exposure;|
| Active Comparator: IVF: overnight exposure<br>Half of a patients' oocytes will be subjected to overnight incubation with sperm (=usual practice). | Other: overnight exposure to sperm for IVF<br>* Oocytes will be exposed for a longer duration (overnight, 16-20h) to progressive motile sperm for insemination<br>* Other names: long exposure;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Normal fertilization rate | presence of two pronuclei per inseminated oocyte | 2 days | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Number of COCs assigned to each group | number of oocytes, per patient, assigned to each exposure group | 2 days | 
| Maturation rate | number of mature oocytes obtained in each arm | 2 days | 
| Fertilization | Number of abnormally fertilized oocytes and failed fertilization | 3 days | 
| Total fertilization failure | Number of cycles with complete fertilization failure (in one arm or in both arms) | 3 days | 
| Embryo quality on day 3 | defined by the Istanbul consensus (Alpha Scientists, 2011) The number of blastomeres and their division pattern, fragmentation, presence of compaction, vacuoles, granulation and nuclei will divide the embryos into 3 categories: good, fair or poor | 5 days | 
| Embryo quality on day 5 | defined by a modified model of Gardner and Schoolcraft,1999 | 8 days | 
| blastulation rate I | number of embryos reaching at least the BL1 stage/ number of oocytes assigned to that group: defined by a 1 (yes, the embryo is blastulating) or 0 (no, the embryo is not blastulating) | 8 days | 
| blastulation rate II | number of embryos reaching at least the BL1 stage/ number of mature oocytes in that group: defined by a 1 (yes, the embryo is blastulating) or 0 (no, the embryo is not blastulating) | 8 days | 
| Utilization rate | number of embryos that can be used for the patient (=number of embryos that can be biopsied)/number of oocytes assigned to the group | 8 days | 
| Day of trophectoderm biopsy | Day at which the blastocyst reaches sufficient expansion to perform biopsy of trophectoderm cells | 8 days | 
| Euploid rate | Number of genetically normal embryos per total number of blastocysts biopsied, stratified per day of biopsy | 8 days | 
| Morphokinetic parameters | As embryos will be monitored in a time lapse imaging system, morphokinetic parameters will be compared between both exposure arms: o t2: the timing to two cells t3: the timing to three cells t4: the timing to four cells t5: the timing to five cells t8: the timing to eight cells SC: starting to compact M: the timing to morula SB: starting to blastulate B: reaching a BL3 according to Gardner and Schoolcraft, 1999 F: reaching a BL4 according to Gardner and Schoolcraft, 1999 cc2: t3-t2 s2: t4-t3 | 10 days | 
| Pregnancy outcome | dichotomous variable defined by the presence of βhCG test of > 15IU 12-15 days after embryo transfer | 60 days | 
| Biochemical pregnancy | dichotomous variable defined as a pregnancy in which the hCG levels start do decrease after 1 week | 70 days | 
| Implantation rate | defined by the number of gestational sacs/number of embryos transferred • Clinical pregnancy (yes or no) defined by the ultrasonographic visualization of one or more gestational sacs, including ectopic pregnancies | 80 days | 
| Clinical pregnancy Fetal Heart beat positive | dichotomous variable defined by the ultrasonographic visualization of one or more gestational sacs, with fetal heart beat at 7 weeks. | 80 days |