Oocyte Activation: Revised or Initial AGT Technique, Which is Better?! A Sibling-oocytes-split Design

Study Overview
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Brief Summary
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This study will be conducted to compare two different Assisted Gamete treatment (AGT) with oocyte-sibling-split design control group : AGT- initial protocol for activating oocytes by culturing oocytes in calcium ionophore after injection for two rounds of 10 minutes at 37 C, 30 minutes after injection. sample group : AGT-revised protocol oocytes will be injected by sperm previously cultured in Ca ionophore and 0.4 pL of calcium ionophore then will be cultured in calcium ionophore after injection for 10 minutes

Detailed Description
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investigators performed ICSI then with one of two activation protocols added : AGT-initial or AGT-revised For the AGT initial protocol, Oocytes will be injected then will be incubated in 10 mM calcium ionophore (Sigma-Aldrich) for two rounds of 10 minutes at 37 C, 30 minutes after ICSI injection, and then will be rinsed and transferred into culture medium. For the AGT-revised protocol, ejaculated spermatozoa will be exposed to calcium ionophore in a drop on the ICSI dish before injection. During the ICSI procedure, spermatozoa will be aspirated individually from the drop containing calcium ionophore and immobilized in a separate polyvinylpyrrolidone drop. Next, approximately 0.4 pL of calcium ionophore will be aspirated into the micropipette and will be injected into the oocyte with the spermatozoa. Post-ICSI oocytes will be then exposed to 50 mM calcium ionophore for 10 minutes at 37 C, and then will be washed and placed in culture medium.

Official Title
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Oocyte Activation: Revised or Initial Activated Gamete Treatment (AGT) Technique, Which is Better?! A Sibling-oocytes-split Design and Case Series of Chemical Pregnancies

Conditions
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Assisted Reproductive Technology

Intervention / Treatment
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* Procedure: Assisted gamete activation


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: couples presented with an extremely poor fertilization (<10%) couples with previous complete fertilization failure after undergoing ICSI cycles At least 4 MII oocytes Severe oligoasthinozoospermia Teratozoospermia testicular aspirated sperm Exclusion Criteria: age >40 years BMI >34 aspirated oocytes < 4 Normozoospermia subfertile patients

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

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Single Group Assignment
Interventional Model Description: single group of patients will be conducted into sibling-oocyte-split design
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: AGT-Initial<br>Oocytes will be injected with spermatozoa and will be incubated in 10 mM calcium ionophore (Sigma-Aldrich) for two rounds of 10 minutes at 37 C, 30 minutes after ICSI injection, and then will be rinsed and transferred into culture medium. | Procedure: Assisted gamete activation<br>* compare two different assisted gamete activation protocols to improve ICSI outcome<br>|
| Experimental: AGT-revised<br>ejaculated spermatozoa will be exposed to calcium ionophore in a drop on the ICSI dish before injection. During the ICSI procedure, spermatozoa will be aspirated individually from the drop containing calcium ionophore and immobilized in a separate PVP drop. Next, approximately 0.4 pL of calcium ionophore will be aspirated into the micropipette and injected into the oocyte with the spermatozoa. Post-ICSI oocytes will be then exposed to 50 mM calcium ionophore for 10 minutes at 37C, and then will be washed and placed in culture medium. | Procedure: Assisted gamete activation<br>* compare two different assisted gamete activation protocols to improve ICSI outcome<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Number of top quality day 3 embryos | number of top quality embryos | 3 days after ICSI procedure day | 
| Number of Top quality day 4 embryos | number of top quality embryos | 4 days after ICSI procedure | 
| Number of Top quality day 5 embryos | number of top quality embryos | 5 days after ICSI procedure | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| chemical pregnancy rate | the number of positive pregnancy cases from overall cases | 15 days after embryo transfer procedure day | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Calcium ionophore, gamete activation, ICSI, male factor