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Effects of calcium ionophore on human oocyte in vitro maturation outcome, fertilization rate and embryo development

Protocol summary

Study aim
Determination of the effects of calcium ionophore on human oocyte in vitro maturation outcome, fertilization rate and embryo development
Design
In this controlled clinical trial, participants are divided into two groups of intervention and control. In this study, immature immature oocytes unable use for ICSI will be used in this study after getting participants consent. Then, after approval by the ethics committee of Shahid Beheshti University of Medical Sciences, patients who undergo ovulation stimulation for ICSI are enrolled in the study. Sample size includes 210 oocytes and 150 embryos that obtained from study participants.
Settings and conduct
Germinal vesicle stage (GV) oocytes enter the two groups: intervention group (Treatment with calcium ionophore) and control group (Group without calcium ionophore) After 24-28 hours, in IVM-derived MII oocytes, nuclear maturation rate , cytoplasm maturation rate, cytoplasm oxidative stress level, meiotic spindle structure, apoptosis rate and MPF and MAPK genes expression will be evaluated. Subsequently, MII oocytes from IVM in the treatment group with ionophosphorus calcium and the control group will be under microinjection with sperm. fertilization rate, cleavage rate and embryo quality will be evaluated .
Participants/Inclusion and exclusion criteria
Inclusion criteria: Approval of the Ethics Committee of Shahid Beheshti University of Medical Sciences, women age between 20-40 years. Sperms have a normal parameter according to WHO criteria Exit criteria:Endometriosis, Polycystic ovarian syndrome, Chromosomal abnormalities
Intervention groups
1. Treatment with calcium ionophore: Each oocytes is placed in a 30-ml drop of ionophore calcium A23187 for 15 minutes, then washed in two 30-microliter drops of the culture medium. 2. Group without calcium ionophore (control)
Main outcome variables
Glutathione level, expression of MPF and MAPK, meiotic spindle structure, ROS level, expression of BAX, BCL2 and caspase-3 genes, fertilization rate, embryo quality, cleavage rate

General information

Reason for update
Acronym
IRCT registration information
IRCT registration number: IRCT20140707018381N4
Registration date: 2018-08-03, 1397/05/12
Registration timing: registered_while_recruiting

Last update: 2018-08-03, 1397/05/12
Update count: 0
Registration date
2018-08-03, 1397/05/12
Registrant information
Name
Ahmad Hosseini
Name of organization / entity
Shahid Beheshty University of Medical Science
Country
Iran (Islamic Republic of)
Phone
+98 21 2235 9153
Email address
ahmad-hosseini@sbmu.ac.ir
Recruitment status
Recruitment complete
Funding source
Expected recruitment start date
2017-10-23, 1396/08/01
Expected recruitment end date
2018-10-23, 1397/08/01
Actual recruitment start date
empty
Actual recruitment end date
empty
Trial completion date
empty
Scientific title
Effects of calcium ionophore on human oocyte in vitro maturation outcome, fertilization rate and embryo development
Public title
Effects of calcium ionophore on human oocyte in vitro maturation outcome
Purpose
Treatment
Inclusion/Exclusion criteria
Inclusion criteria:
Approval of the Ethics Committee of Shahid Beheshti University of Medical Sciences The age of a woman is between 20-40 years old Sperm has a normal parameter based on WHO criteria
Exclusion criteria:
Endometriosis Poly cystic ovary syndrome Genetic disorders
Age
From 20 years old to 40 years old
Gender
Both
Phase
3
Groups that have been masked
No information
Sample size
Target sample size: 150
Randomization (investigator's opinion)
Randomized
Randomization description
The sampling method will be two-stage sampling methods. In the first stage, non-probability sampling method is used and in the second stage, random allocation is performed using permuted block randomization between the two intervention and control groups.
Blinding (investigator's opinion)
Not blinded
Blinding description
Placebo
Not used
Assignment
Parallel
Other design features

Secondary Ids

empty

Ethics committees

1

Ethics committee
Name of ethics committee
Ethics Committee of Shahid Beheshti University of Medical Sciences
Street address
Faculty of medicine, Arabi Ave, Daneshjo street, Velenjak, Tehran
City
Tehran
Province
Tehran
Postal code
1985717443
Approval date
2017-09-28, 1396/07/06
Ethics committee reference number
IR.SBMU.MSP.1396.416

Health conditions studied

1

Description of health condition studied
Infertility
ICD-10 code
N97.9
ICD-10 code description
Female infertility, unspecified

Primary outcomes

1

Description
Oocyte cytoplasmic maturation
Timepoint
After 24-28 hours, the culture was incubated at 37 ° C and 5% CO2.
Method of measurement
Cell Tracker Blue staining for oocyte cytoplasmic maturity

2

Description
Oocyte oxidative stress
Timepoint
After 24-28 hours, the culture was incubated at 37 ° C and 5% CO2.
Method of measurement
Evaluation of the free radical oxygen (ROS) in cytoplasmic by staining with H2DCFDA

3

Description
Meiotic spindle structure
Timepoint
After 24-28 hours, the culture was incubated at 37 ° C and 5% CO2.
Method of measurement
Evaluation of miieotic spindle structure by staining with anti-tubulin antibody

4

Description
Apoptosis rate
Timepoint
After 24-28 hours, the culture was incubated at 37 ° C and 5% CO2.
Method of measurement
Determination of apoptosis rate by evaluating expression bax, bcl2 and caspase-3 gene by Real Time-PCR

5

Description
Nucleation maturation
Timepoint
After 24-28 hours, the culture was incubated at 37 ° C and 5% CO2.
Method of measurement
Determining the oocyte nucleus maturity by evaluating expression of MPF and MAPK by Real Time-PCR

Secondary outcomes

1

Description
Fertilization rate
Timepoint
16-18 hours after ICSI
Method of measurement
Divide the total number of oocytes fertilized to the total oocytes under ICSI

2

Description
Cleavage rate
Timepoint
The third day after ICSI
Method of measurement
Split the total number of cleavage embryos divided into fertilized embryos

3

Description
Embryo quility
Timepoint
The third day after ICSI
Method of measurement
Grading of embryo morphology based on the number and size of the blastomeres and the rate of fragmentation

Intervention groups

1

Description
Intervention group: Treatment group with calcium ionophore: Each oocyte is placed in a 30-ml droplet of ionophore calcium A23187 for 15 minutes, then washed in two drops of 30 μL of the culture medium. The oocytes are then culture in an ecobar for 24-28 hours.
Category
Treatment - Drugs

2

Description
Control group: Group without treatment with ionophore calcium
Category
Treatment - Drugs

Recruitment centers

1

Recruitment center
Name of recruitment center
Taleghani hospital
Full name of responsible person
Elham Fazeli
Street address
Taleghani hospital, Arabi Ave, Daneshjo street, Velenjak, Tehran
City
Tehran
Province
Tehran
Postal code
1985711151
Phone
+98 21 2243 2560
Email
Elhamfazeli15@yahoo.com

Sponsors / Funding sources

1

Sponsor
Name of organization / entity
Shahid Beheshti University of Medical Sciences
Full name of responsible person
Marefat Ghaffari
Street address
Faculty of medicine, Arabi Ave, Daneshjo street, Velenjak, Tehran
City
Tehran
Province
Tehran
Postal code
1985717443
Phone
+98 21 2243 9770
Email
Mghaffarin@yahoo.com
Grant name
Grant code / Reference number
Is the source of funding the same sponsor organization/entity?
Yes
Title of funding source
Shahid Beheshti University of Medical Sciences
Proportion provided by this source
100
Public or private sector
Public
Domestic or foreign origin
Domestic
Category of foreign source of funding
empty
Country of origin
Type of organization providing the funding
Academic

Person responsible for general inquiries

Contact
Name of organization / entity
Shahid Beheshti University of Medical Sciences
Full name of responsible person
Elham Fazeli
Position
PhD candidate
Latest degree
Master
Other areas of specialty/work
Reproductive Biology
Street address
Faculty of medicine, Arabi Ave, Daneshjo street, Velenjak, Tehran
City
Tehran
Province
Tehran
Postal code
1983969411
Phone
+98 13425965
Email
Elhamfazeli15@yahoo.com

Person responsible for scientific inquiries

Contact
Name of organization / entity
Shahid Beheshti University of Medical Sciences
Full name of responsible person
Elham Fazeli
Position
PhD candidate
Latest degree
Master
Other areas of specialty/work
Reproductive Biology
Street address
Faculty of medicine, Arabi Ave, Daneshjo street, Velenjak, Tehran
City
Tehran
Province
Tehran
Postal code
1983969411
Phone
+98 13425965
Email
Elhamfazeli15@yahoo.com

Person responsible for updating data

Contact
Name of organization / entity
Shahid Beheshti University of Medical Sciences
Full name of responsible person
Elham Fazeli
Position
PhD candidate
Latest degree
Master
Other areas of specialty/work
Reproductive Biology
Street address
Shahid Beheshti University of medical science, Arabi Ave, daneshjo st, Velenjak, Tehran
City
Tehran
Province
Tehran
Postal code
1983969411
Phone
+98 13 4259 6507
Email
Elhamfazeli15@yahoo.com

Sharing plan

Deidentified Individual Participant Data Set (IPD)
Yes - There is a plan to make this available
Study Protocol
Yes - There is a plan to make this available
Statistical Analysis Plan
Yes - There is a plan to make this available
Informed Consent Form
Yes - There is a plan to make this available
Clinical Study Report
Yes - There is a plan to make this available
Analytic Code
Yes - There is a plan to make this available
Data Dictionary
Not applicable
Title and more details about the data/document
All interventions in the patient's file will be recorded and archived in the Taleghani hospital. All data obtained from this study will be published with regard to the privacy and confidentiality right of the the information of research participants.
When the data will become available and for how long
Get started 6 months after publishing study results
To whom data/document is available
The findings of this study will be accessible to all individuals.
Under which criteria data/document could be used
To improve pregnancy outcomes in infertility clinics
From where data/document is obtainable
Elham Fazeli Email: Elhamfazeli15@yahoo.com Tel:00989112448530
What processes are involved for a request to access data/document
Data that release in the published article will be available to other people according to the journal protocol that published article (free or purchase). In order to access data that not published in the paper, it is required to obtain the correspond author's approval and then approval of the research deputy of Shahid Beheshti University of Medical Sciences is obligatory.
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