Oocyte in vitro maturation (IVM) is a minimal-stimulation ART with reduced hormone-related side effects and risks for the patients. However, the approach is not widely used because of an efficiency gap compared to conventional ART. In order to further optimize and adapt the CAPA-IVM system in the IVM clinic, this pilot study aims to check the feasibility of applying a single COC CAPA-IVM strategy versus the group COC culture CAPA-IVM
Oocyte in vitro maturation (IVM) is a minimal-stimulation ART with reduced hormone-related side effects and risks for the patients. However, the approach is not widely used because of an efficiency gap compared to conventional ART. Oocytes retrieved for IVM procedures derive from a heterogeneous pool with variable cellular and molecular characteristics that indicate its immature status. Thus, in vitro systems that permit and enhance acquisition and synchronization of meiotic competence (ability to resume meiosis in response to an ovulatory stimulus) and developmental competence (ability to be fertilized and support early embryo development) before the meiotic trigger are crucial for the optimization of human IVM systems. A novel two-step IVM culture system (named CAPA-IVM) involving a pre-maturation culture with C-type natriuretic peptide (CNP) and a maturation step in presence of Amphiregulin (AREG), both more physiological compounds improving oocyte competence, have been introduced in previous clinical studies. So far these pilot studies proved to increase the rates of oocyte maturation, good quality embryos on day 3, good quality blastocyst, and as a result a higher embryo yield. CAPA-IVM blastocysts have shown similar rates of methylation and gene expression at gDMRs compared to COS embryos; and the expression of ma-jor epigenetic regulators was similar between both groups. Furthermore, an improvement in pregnancy rates strengthens the clinical relevance of the use of CAPA-IVM strategy. In order to further optimize and adapt the CAPA-IVM system in the IVM clinic, this pilot study aims to check the feasibility of applying a single COC CAPA-IVM strategy versus the group COC culture CAPA-IVM. A single COC culture would permit to perform a non-invasive molecular analysis per matured oocyte, in order to identify quality genes ex-pressed in cumulus cells post-IVM (cumulus biomarkers), which could be subsequently used to identify the embryo(s) with highest potential of implantation.
Study Type
INTERVENTIONAL
Allocation
NON_RANDOMIZED
Purpose
TREATMENT
Masking
NONE
Enrollment
15
Cumulus-oocyte complexes (COC) will be cultured in CAPA-IVM standard conditions: 24 hrs capacitation followed by 30h maturation. The first group will be culture in pools (group culture). The second group will be culture individually in 20µl droplets.
My Duc Hospital
Ho Chi Minh City, Vietnam
Number of good quality embryos
Number of good quality Day 3 embryos obtained
Time frame: At least 3 days after intra-cytoplasmic sperm injection
Maturation rate
Percentage of mature oocytes by 2 types of COC culture
Time frame: Two days after oocytes pick-up
Fertilization rate
Percentage of fertilized oocytes by 2 types of COC culture
Time frame: 16-18 hours after intra-cytoplasmic sperm injection
Cleavage rate
Percentage of day-3 embryos over fertilized oocytes by 2 types of COC culture
Time frame: At least 3 days after intra-cytoplasmic sperm injection
Expansion rate
Percentage of cumulus-oocyte complexes expanded after culture by 2 types of COC culture
Time frame: After at least 30 hours of maturation culture
The relative expression ratio ( R ) of human cumulus cell genes
Cumulus cells will be collected, cDNA synthesis after mRNA purification, relative quantification PCR for detecting gene expression
Time frame: cumulus cells will be collected after at least 30 hours of maturation culture, storaged at -80oC until RNA purification
Positive pregnancy test
Serum human chorionic gonadotropin level greater than 25 mIU/mL
Time frame: at 2 weeks after the embryo placement after the completion of the first transfer
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Clinical pregnancy
at least one gestational sac on ultrasound at 7 weeks' gestation with the detection of heart beat activity
Time frame: 5 weeks after embryo placement after the completion of the first transfer
Ongoing pregnancy
Ongoing pregnancy is defined as pregnancy with detectable heart rate at 12 weeks' gestation or beyond, after the completion of the first transfer
Time frame: At 12 weeks' gestation
Implantation rate
as the number of gestational sacs per number of embryos transferred
Time frame: 3 weeks after embryo transferred after the completion of the first transfer
Multiple pregnancy
Defined as presence of more than one sac at early pregnancy ultrasound (6-8 weeks gestation)
Time frame: 5 weeks after embryo placement after the completion of the first transfer
Multiple delivery
Birth of more than one baby beyond 24 weeks
Time frame: At 24 weeks' gestation
Miscarriage
pregnancy loss at \< 24 weeks
Time frame: at 24 weeks of gestation after the completion of the first transfer
Live birth
defined as at least one newborn after 24 weeks of gestation and exhibiting any sign of life; twins were counted as a single birth
Time frame: At 24 weeks of gestation