Cytokines Supplementation to Culture Media and ICSI Outcome

Study Overview
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Brief Summary
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A randomized clinical trial to evaluate the effect of cytokine supplementation in embryo culture medium with various protein concentrations aiming to evaluate the effects of cytokines (CYK) integration into the human in vitro culture medium with high and low human serum albumin (HSA) concentrations on live birth rates after intracytoplasmic sperm injection (ICSI).

Detailed Description
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Oocytes were injected, and embryos cultured and transferred in three supplemented culture media. A- A single step culture medium (SSCM; Global, Life Global)+5mg/ml (10% Vol/Vol), life global protein, without supplementation of CYK (control n=200), B- A single step culture medium (SSCM; Global, Life Global)+5mg/ml (10% Vol/Vol), life global protein, with 2ng/mL GM-CSF (granulocyte macrophage colony stimulating factor) (G5035 Sigma), 5ng/mL HB-EGF (Heparine Binding Epidermal Growth Factor )(E4643 Sigma),and 5ng/mL LIF (Lukemia Inhbitory Factor) (SRP9001 Sigma)(n=200). C- A single step culture medium (SSCM; Global, Life Global) +2mg/ml (5% Vol/Vol) life global protein, with 2ng/mL GM-CSF (G5035 Sigma), 5ng/mL HB-EGF (E4643 Sigma), and 5ng/mL LIF (SRP9001 Sigma).(n=200) Setting :- - Mansoura Integrated Fertility

Official Title
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Impact of Cytokine Supplementation With Various Protein Concentrations Into Human in Vitro Culture Medium on ICSI Cycle Outcome

Conditions
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ICSI

Intervention / Treatment
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* Combination Product: A single step culture medium


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Patients who fitted the medical definition of infertility One year of unprotected intercourse but not pregnant. Exclusion Criteria: History of ovarian or adnexal surgery. Suspicious findings of ovarian malignancy. Presence of endocrine disorders such as diabetes mellitus, hyper-prolactinemia, thyroid dysfunction, congenital adrenal hyperplasia, Cushing's syndrome, and adrenal insufficiency. Presence of Globozoospermia (100% round-headed spermatozoa)

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 38 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: Multicenter, randomized, double-blinded and prospective study
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: HSA<br>A single step culture medium (SSCM; Global, Life Global)+5mg/ml (10% Vol/Vol) life global protein, | Combination Product: A single step culture medium<br>* A single step culture medium (SSCM; Global, Life Global)+5mg/ml life global protein, without supplementation of CYK<br>|
| Experimental: CYK+ high HSA<br>A single step culture medium (SSCM; Global, Life Global)+5mg/ml (10% Vol/Vol) life global protein, supplemented with 2ng/mL GM-CSF (G5035 Sigma), 5ng/mL HB-EGF (E4643 Sigma),and 5ng/mL LIF (SRP9001 Sigma). | Combination Product: A single step culture medium<br>* A single step culture medium (SSCM; Global, Life Global)+5mg/ml life global protein, without supplementation of CYK<br>|
| Active Comparator: CYK+low HSA<br>A single step culture medium (SSCM; Global, Life Global) +2mg/ml (5% Vol/Vol) life global protein, supplemented with 2ng/mL GM-CSF (G5035 Sigma), 5ng/mL HB-EGF (E4643 Sigma), and 5ng/mL LIF (SRP9001 Sigma). | Combination Product: A single step culture medium<br>* A single step culture medium (SSCM; Global, Life Global)+5mg/ml life global protein, without supplementation of CYK<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| live birth rate | the number of deliveries that resulted in at least one live born baby (>20 weeks gestational age or 500 gm birth weight ) expressed per 100 embryo transfer, | 9 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Fertilization rate | Fertilization rate (fertilized oocytes with two pronuclei/ number of MII oocytes) ×100 % | 4 weeks | 
| Top quality day 3 embryos | embryos with seven or eight blastomeres of stage-proper sizes, and <10% fragmentation by volume | 4 weeks | 
| Blastocyst formation rate | (number of blastocysts/number of cultured embryos for blastocyst formation) ×100% | 4 weeks | 
| Top blastocyst rate | (number of high quality blastocysts/number of formed blastocysts) ×100% | 4 weeks | 
| Available embryo rate | number of transferred and frozen embryos/number of zygotes (2 pronuclei) ×100% | 4 weeks | 
| clinical pregnancy rate | number of cycles with clinical pregnancy/number of transferred cycles) ×100% | 8 weeks | 
| Biochemical pregnancy rate | number of cycles with biochemical pregnancy/number of Cycles with positive HCG) ×100%. | 6 weeks | 
| Implantation rate | number of implanted embryos/number of transferred embryos) ×100% | 8 weeks | 
| Early embryo loss rat | number of early spontaneously aborted embryos/number of transferred embryos) × 100%. | 16 weeks | 
| ongoing pregnancy rate | number of clinical pregnancies after 12 weeks divided by number of embryo transfers multiplied by 100 | 16 weeks |