Methods for Fertility Preservation: Impact of Vitrification on in Vitro Matured Oocytes
Overview
- Phase
- Not Applicable
- Intervention
- Not specified
- Conditions
- Fertility
- Sponsor
- University Hospital, Clermont-Ferrand
- Enrollment
- 240
- Locations
- 1
- Primary Endpoint
- The embryonic development kinetics
- Last Updated
- 7 years ago
Overview
Brief Summary
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.
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.
Investigators
Eligibility Criteria
Inclusion Criteria
- •ICSI treatment
- •Immature oocytes
- •Without ovulation pathologies
Exclusion Criteria
- •Polykistic ovarian syndrome
- •Endometriosis
- •Ovulatory disease
Outcomes
Primary Outcomes
The embryonic development kinetics
Time Frame: 6 days after ICSI and through study completion
from the records obtained with Time Lapse Primovision, investigator will be able to determine the precise times of embryonic development after in vitro maturation.
Secondary Outcomes
- Analysis of actin and tubulin cytoskeleton and spindle organization in mature ovocytes (Metaphase II)(01/01/2019 - 31/12/2019)
- Analysis of chromosome segregation during the first meiotic division(01/01/2019 - 31/12/2019)
- Analysis of cortical granules distribution in mature (Metaphase II) oocytes.(01/01/2019 - 31/12/2019)
- Analysis of maternal factor stabilities.(01/01/2019 - 31/12/2019)