Single vs. Group CAPA-IVM Culture of Cumulus-oocyte Complexes

Effect of Single vs. Group CAPA-IVM Culture of Human Cumulus-oocyte Complexes From Small Antral Follicles in a SIBLING Oocyte Study Design

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

Study Overview

• Status: Completed
• Conditions: PCOS; IVF; IVM
• Intervention / Treatment: Other: The way of Cumulus-oocyte complexes (COC) culture

Detailed Description

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
• Enrollment (Actual): 15
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Vietnam
  • Ho Chi Minh City, Vietnam
    • My Duc Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 37 years (Adult)
• Accepts Healthy Volunteers: Yes

Description

Inclusion Criteria:

• Having polycystic ovarian morphology: at least 25 follicles (2-9 mm) throughout the whole ovary and/or increased ovarian volume (>10ml) (it is sufficient that 1 ovary fits these criteria)
• No major uterine or ovarian abnormalities
• Having at least 15 follicles on the OPU day
• Patients consent to participate in the study

Exclusion Criteria:

• High (>grade 2) grade endometriosis
• Cases with extremely poor sperm (serve OAT: density <1 million, mobility <10% and sperm from testicular surgery)

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Non-Randomized
• Interventional Model: Parallel Assignment
• Masking: None (Open Label)

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Single Culture; Transfer the COCs cultured individually in Capacitation medium to the individual washing droplets from the "washing dish" containing "Maturation Medium" and wash them thoroughly. Then transfer COCs one by one to the "Culture dish" with "Maturation Medium".	Other: The way of Cumulus-oocyte complexes (COC) culture; 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.
Experimental: Group Culture; Transfer half of the COCs (5-10 at a time) from the Capacitation culture dish to the "washing dish" containing "Maturation Medium (Group Culture)" by using an Eppendorf micropipette and wash them thoroughly (load pipette tips with 5µl, max. 10µl). Then transfer COCs to the "IVM dish" with "Maturation Medium (Group Culture)".	Other: The way of Cumulus-oocyte complexes (COC) culture; 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.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Number of good quality embryos	Number of good quality Day 3 embryos obtained	At least 3 days after intra-cytoplasmic sperm injection

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Multiple delivery	Birth of more than one baby beyond 24 weeks	At 24 weeks' gestation
Maturation rate	Percentage of mature oocytes by 2 types of COC culture	Two days after oocytes pick-up
Fertilization rate	Percentage of fertilized oocytes by 2 types of COC culture	16-18 hours after intra-cytoplasmic sperm injection
Cleavage rate	Percentage of day-3 embryos over fertilized oocytes by 2 types of COC culture	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	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	cumulus cells will be collected after at least 30 hours of maturation culture, storaged at -80oC until RNA purification
Clinical pregnancy	at least one gestational sac on ultrasound at 7 weeks' gestation with the detection of heart beat activity	5 weeks after embryo placement after the completion of the first transfer
Implantation rate	as the number of gestational sacs per number of embryos transferred	3 weeks after embryo transferred 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	At 12 weeks' gestation
Miscarriage	pregnancy loss at < 24 weeks	at 24 weeks of gestation 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)	5 weeks after embryo placement after the completion of the first transfer
Positive pregnancy test	Serum human chorionic gonadotropin level greater than 25 mIU/mL	at 2 weeks after the embryo placement 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	At 24 weeks of gestation

Collaborators and Investigators

• Sponsor: Mỹ Đức Hospital
• Investigators: Principal Investigator: Tuong M Ho, MD, MCE, Hope Research Center

Publications and helpful links

General Publications

• Sanchez F, Le AH, Ho VNA, Romero S, Van Ranst H, De Vos M, Gilchrist RB, Ho TM, Vuong LN, Smitz J. Biphasic in vitro maturation (CAPA-IVM) specifically improves the developmental capacity of oocytes from small antral follicles. J Assist Reprod Genet. 2019 Oct;36(10):2135-2144. doi: 10.1007/s10815-019-01551-5. Epub 2019 Aug 9.
• Vuong LN, Le AH, Ho VNA, Pham TD, Sanchez F, Romero S, De Vos M, Ho TM, Gilchrist RB, Smitz J. Live births after oocyte in vitro maturation with a prematuration step in women with polycystic ovary syndrome. J Assist Reprod Genet. 2020 Feb;37(2):347-357. doi: 10.1007/s10815-019-01677-6. Epub 2020 Jan 4.
• Sanchez F, Romero S, De Vos M, Verheyen G, Smitz J. Human cumulus-enclosed germinal vesicle oocytes from early antral follicles reveal heterogeneous cellular and molecular features associated with in vitro maturation capacity. Hum Reprod. 2015 Jun;30(6):1396-409. doi: 10.1093/humrep/dev083. Epub 2015 Apr 22.
• Saenz-de-Juano MD, Ivanova E, Billooye K, Herta AC, Smitz J, Kelsey G, Anckaert E. Genome-wide assessment of DNA methylation in mouse oocytes reveals effects associated with in vitro growth, superovulation, and sexual maturity. Clin Epigenetics. 2019 Dec 19;11(1):197. doi: 10.1186/s13148-019-0794-y. Erratum In: Clin Epigenetics. 2020 Jan 27;12(1):18.
• Sanchez F, Lolicato F, Romero S, De Vos M, Van Ranst H, Verheyen G, Anckaert E, Smitz JEJ. An improved IVM method for cumulus-oocyte complexes from small follicles in polycystic ovary syndrome patients enhances oocyte competence and embryo yield. Hum Reprod. 2017 Oct 1;32(10):2056-2068. doi: 10.1093/humrep/dex262.

Study record dates

Study Major Dates

• Study Start (Actual): October 1, 2020
• Primary Completion (Actual): November 30, 2020
• Study Completion (Actual): October 24, 2021

Study Registration Dates

• First Submitted: September 5, 2020
• First Submitted That Met QC Criteria: September 23, 2020
• First Posted (Actual): September 24, 2020

Study Record Updates

• Last Update Posted (Actual): July 25, 2023
• Last Update Submitted That Met QC Criteria: July 23, 2023
• Last Verified: July 1, 2023

More Information

Terms related to this study

• Keywords: Group culture; CAPA-IVM; Single culture
• Other Study ID Numbers: 13/20/DD-BVMD

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

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