Methods for Fertility Preservation: Impact of Vitrification on in Vitro Matured Oocytes (OVOMIV)

During the last decades, there was an improvement of the cancer treatments of the woman and the teenagers. Therefore higher survival rate is described. However, cancer treatments can alter the reproduction functions and reduce considerably the window of the fertility to the adulthood. Therefore, it is recommended to proceed to a fertility preservation by oocytes vitrification when it is possible. The vitrification is a freezing technique allowing high survival rate and similar results by assisted reproductive technologies compared with the use of fresh oocytes. An innovative method of automated vitrification was recently developed. The usual protocol consist to vitrify mature oocytes. However, this strategy cannot be used for hormone -sensitive cancer or when ovarian stimulation is not possible. In these situations, immature oocytes can be collected. It is also necessary to realize an in vitro maturation step for a use by assisted reproductive technology.

According to the recent data of the literature, it remains unclear whether the vitrification of ovocytes must be performed before or after in vitro maturation (IVM). Therefore the aim of this study is to study the impact on structure and functions of ovocytes when vitrification is performed before or after IVM. The vitrification will be performed by a semi-automatic method which is an innovative method.

Study Overview

• Status: Unknown
• Conditions: Cancer; Fertility
• Intervention / Treatment: Other: Gavi , Merck® (automated vitrification instrument); Other: Vitrification

Detailed Description

To perform this study, investigator will compare three groups. Group 1: immature ovocytes vitrified before IVM; Group 2: immature oocytes vitrified after IVM; Group3: fresh immature oocytes treated by IVM (without vitrification, control group).

The immature oocytes provide from ICSI patients. In routine these oocytes (germinal vesicle) are normally destroyed because they cannot be used for injection. The women will give an informed and written consent. Inclusion criteria are women less 37 years without dysovulation.

The vitrification will be performed with the semi-automatic method (Gavi, Merck). The kinetic and maturation rate will be analysed by time lapse (Primovision, Vitrolife) In the mature oocytes, the actin and tubulin cytoskeleton, the spindle organization and the cortical granules will be studied by immunofluorescence and 3D confocal microscopy. The expression of maternal factors transcription will be analyzed by RT-PCR. The ploidy will be analysed by multiFISH and/or CGH array.

• Study Type: Observational
• Enrollment (Anticipated): 240

Contacts and Locations

Study Locations

• France
  • Clermont-Ferrand, France, 63003
    • CHU Clermont-Ferrand

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 37 years (ADULT)
• Accepts Healthy Volunteers: No
• Genders Eligible for Study: Female

• Sampling Method: Non-Probability Sample

Study Population

women

Description

Inclusion Criteria:

• ICSI treatment
• Immature oocytes
• Without ovulation pathologies

Exclusion Criteria:

• Polykistic ovarian syndrome
• Endometriosis
• Ovulatory disease

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Immature oocytes vitrified before in vitro maturation; Immature oocytes will be vitrified using closed system vitrification with a semiautomatic method. After warming, a maturation culture will be performed during 36 hours	Other: Gavi , Merck® (automated vitrification instrument); Gavi, Merck ® permits semi-automated vitrification with closed system.; Other: Vitrification; Group 1: immature ovocytes vitrified before IVM; Group 2: immature oocytes vitrified after IVM; Group3: fresh immature oocytes treated by IVM (without vitrification, control group).
Immature oocytes vitrified after in vitro maturation; A maturation culture of immature oocytes will be performed in vitro during 36 hours. After IVM, mature oocytes will be vitrify in closed system by a semi-automatic method.	Other: Gavi , Merck® (automated vitrification instrument); Gavi, Merck ® permits semi-automated vitrification with closed system.; Other: Vitrification; Group 1: immature ovocytes vitrified before IVM; Group 2: immature oocytes vitrified after IVM; Group3: fresh immature oocytes treated by IVM (without vitrification, control group).
Fresh oocytes; The culture of immature oocytes will be performed during 36 hours

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
The embryonic development kinetics	from the records obtained with Time Lapse Primovision, investigator will be able to determine the precise times of embryonic development after in vitro maturation.	6 days after ICSI and through study completion

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Analysis of actin and tubulin cytoskeleton and spindle organization in mature ovocytes (Metaphase II)	Metaphase-II stage oocytes will be used for Immuno-Fluorescence experiments to stain actin, tubulin and chromosomes. Oocytes will be imaged using confocal microscope to perform high resolution imaging and quantitative image analysis. The length, position and orientation of the second meiotic spindle will be analysed. The actin network and chromosomes will be analysed quantitatively. All measurements will be compared with fresh matured (Metaphase-II) oocytes used as a control group.	01/01/2019 - 31/12/2019
Analysis of chromosome segregation during the first meiotic division	A multi Fluorescence in Situ Hybridization and/or a CGH array will be performed to measure the chromosome segregation after vitrification	01/01/2019 - 31/12/2019
Analysis of cortical granules distribution in mature (Metaphase II) oocytes.	A staining with Lectin will be used to mark cortical granules of matured oocytes to observe whether the protocol has an impact on their spatial distribution. To analyse this staining, investigator will use quantitative image analysis method.	01/01/2019 - 31/12/2019
Analysis of maternal factor stabilities.	Maternal factors stored in the oocyte cytoplasm during oogenesis as proteins and transcripts are essential for the early embryonic development. Investigator will perform Reverse Transcription combined with Real Time PCR to quantify transcript amounts of candidates genes selected from human oocytes databases.	01/01/2019 - 31/12/2019

Collaborators and Investigators

• Sponsor: University Hospital, Clermont-Ferrand

Study record dates

Study Major Dates

• Study Start (ANTICIPATED): September 20, 2018
• Primary Completion (ANTICIPATED): December 31, 2019
• Study Completion (ANTICIPATED): July 31, 2020

Study Registration Dates

• First Submitted: June 5, 2018
• First Submitted That Met QC Criteria: September 20, 2018
• First Posted (ACTUAL): September 21, 2018

Study Record Updates

• Last Update Posted (ACTUAL): September 21, 2018
• Last Update Submitted That Met QC Criteria: September 20, 2018
• Last Verified: September 1, 2018

More Information

Terms related to this study

• Keywords: Vitrification; In vitro maturation; time lapse; High resolution imaging; quantitative molecular analysis
• Other Study ID Numbers: CHU-393

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No