Scanning the Meiotic Spindle in Assisted Reproductive Techniques to Assess Oocyte Quality

NCT06539585 | Active Not Recruiting

• 1 other identifier: MW24-08-00048
• Study Type: interventional
• Target: 164
• Locations: 1 country
• Sites: 1

Brief Summary

The assisted reproduction success rate is affected by several factors including the age of the women, oocyte quality and maturation state, as well as sperm quality. Imaging of the meiotic spindle may be crucial for determining the oocyte maturation and the optimal time of oocyte fertilization by intracytoplasmic sperm injection (ICSI). A new accurate and non-invasive method for selecting quality maturated oocytes based on meiotic spindle imaging is for women over 35 years of age will be introduced. The evaluation of efficiency using meiotic spindle visibility in polarized light and its relative position to the polar body as indicator of oocyte maturity will be monitored and the optimal time for ICSI will be defined.

Conditions

Infertility; Infertility, Male; Infertility, Female

Keywords

infertility; human oocyte; IVF; meiotic spindle; ICSI; sperm count

Outcome Measures

Primary Outcomes (1)

• Clinical pregnancy rate of patients whose ICSI time was established according to MS state

  The percentage of all attempts that leads to pregnancy of patients whose ICSI fertilization was done according MS status. Meiotic spindle visibility in polarized light and its relative position to the polar body as indicator of oocyte maturity will be monitored and the optimal time for ICSI will be defined.

  6-8 weeks

Secondary Outcomes (1)

• clinical pregnancy rate of patients whose ICSI time was not established according to measured MS state

  6-8 weeks

Other Outcomes (1)

• Clinical pregnancy rate of patients with standard ICSI time

  6-8 weeks

Study Arms (3)

Standard ICSI without meiotic spindle imaging.

ACTIVE COMPARATOR

A control group of patients who underwent a standard IVF cycle without microscopy of the meiotic spindle. In this control group of patients, ICSI fertilization will be performed 5-6 hours after oocyte collection. ICSI will be performed according to standard protocol \[6\]. The oocytes will be cultivated individually, and their order preserved, so that the other outcomes (data from timelapse, clinical results) can be associated with individual oocytes. Fertilization after ICSI will be defined as the presence of two pronuclei and 2 polar body. Embryos will be cultivated for 122-144 h.

Procedure: ICSI

Meiotic spindle imaging followed by standard ICSI.

ACTIVE COMPARATOR

MS evaluation will take place in pre-prepared glass-bottomed dishes with about 5μl medium with the HEPES buffer covered with paraffin oil, put to heat one hour before use. The oocyte will be rotated using a needle so that PB and MS are both well visible, and photographs will be taken using optical microscope. The MS status and the angle (α) between MS and PB will be obtained 3-4 hours after oocyte pick-up (OPU). At the same time, the polarized-light microscopy image will be acquired (polarized light microscopy at ×100 magnification. ICSI will be performed typically 2-3 h after the polarization microscopy evaluation, i.e., 5-6 h after OPU. ICSI will be performed according to standard protocol \[6\]. Fertilization after ICSI will be defined as the presence of two pronuclei and 2 polar body. Embryos will be cultivated for 122-144 h.

Procedure: Meiotic spindle status observation in polarized light; Procedure: ICSI

MS imaging and ICSI fertilization according MS status.

ACTIVE COMPARATOR

Oocytes with MS evaluation will be fertilized according to MS status either 5-6 hours after ovum pick-up (OPU) or 7-8 hours after OPU. MS evaluation will take place in pre-prepared glass-bottomed dishes. The MS status and the angle (α) between MS and PB will be obtained 3-4 hours after oocyte pick-up (OPU). At the same time, the polarized-light microscopy image will be acquired (polarized light microscopy at ×100 magnification. For oocytes with PB/MS in close proximity (angle between PB and MS \< 5◦) or MS not visible ICSI will be performed 4-5 h after the polarization microscopy evaluation, i.e., 7-8 h after OPU (these oocytes are supposed to be not fully mature). For oocytes with MS clearly visible and PB/MS not in close proximity (angle between PB and MS \> 5◦) ICSI will be performed typically 2-3 h after the polarization microscopy evaluation, i.e., 5-6 h after OPU. ICSI will be performed according to standard protocol \[6\].

Procedure: Meiotic spindle status observation in polarized light; Procedure: ICSI

Interventions

Meiotic spindle status observation in polarized light PROCEDURE

In patients older than 35 years and younger than 40 years, we will use a microscope with a polarizing filter to evaluate the position of meiotic spindles and polar bodies in oocytes collected from patients who were indicated for IVF and ICSI. Using an optical microscope with a Nikon CEE GmbH polarizing filter, the angle between PB and MS together with MS visibility will be determined.

MS imaging and ICSI fertilization according MS status.; Meiotic spindle imaging followed by standard ICSI.

ICSI PROCEDURE

ICSI will be performed according to standard protocol using ICSI/holding micropipettes (#002-5-30/#001-120-30, Microtech IVF, Czech Republic), polyvinylpyrrolidone (ICSI™, Vitrolife, Sweden), and Eppendorf (Hamburg, Germany) micromanipulation system equipped with thermoplate (Tokaohit, Japan). The oocytes will be cultivated individually, and their order preserved, so that the other outcomes (data from timelapse, clinical results) can be associated with individual oocytes. Oocytes will be denuded (HYASE-10X™, Vitrolife, Sweden) after OPU, and the maturation stage will be examined.

MS imaging and ICSI fertilization according MS status.; Meiotic spindle imaging followed by standard ICSI.; Standard ICSI without meiotic spindle imaging.

Eligibility Criteria

• Age: 35 Years - 40 Years
• Healthy Volunteers: No
• Age Groups: Adult (18-64)

You may qualify if:

• Women who underwent ICSI;
• Spermiogram containing at least 0.5 mil/ml sperms;
• Morphlogy: normal morphology \>1%;
• Stimulated cycles

You may not qualify if:

• Age under 35 years or over 40;
• Native cycles;
• Severe uterine abnormalities (submucosal fibroid, fibroid ≥5cm, uterine septum, endometrial polyps, uterus duplex)

Sponsors & Collaborators

• Charles University, Czech Republic: lead
• Czech Academy of Sciences: collaborator
• Czech Technical University in Prague: collaborator
• General University Hospital, Prague: collaborator

Study Sites (1)

General University Hospital in Prague

Prague, 128 08, Czechia

Location

Related Publications (7)

• van Loendersloot LL, van Wely M, Limpens J, Bossuyt PM, Repping S, van der Veen F. Predictive factors in in vitro fertilization (IVF): a systematic review and meta-analysis. Hum Reprod Update. 2010 Nov-Dec;16(6):577-89. doi: 10.1093/humupd/dmq015. Epub 2010 Jun 25.

  PMID: 20581128 BACKGROUND

• Wu B, Shi J, Zhao W, Lu S, Silva M, Gelety TJ. Understanding reproducibility of human IVF traits to predict next IVF cycle outcome. J Assist Reprod Genet. 2014 Oct;31(10):1323-30. doi: 10.1007/s10815-014-0288-y. Epub 2014 Aug 15.

  PMID: 25119191 BACKGROUND

• Hanevik HI, Hessen DO. IVF and human evolution. Hum Reprod Update. 2022 Jun 30;28(4):457-479. doi: 10.1093/humupd/dmac014.

  PMID: 35355060 BACKGROUND

• Rienzi L, Ubaldi F, Martinez F, Iacobelli M, Minasi MG, Ferrero S, Tesarik J, Greco E. Relationship between meiotic spindle location with regard to the polar body position and oocyte developmental potential after ICSI. Hum Reprod. 2003 Jun;18(6):1289-93. doi: 10.1093/humrep/deg274.

  PMID: 12773461 BACKGROUND

• Innocenti F, Fiorentino G, Cimadomo D, Soscia D, Garagna S, Rienzi L, Ubaldi FM, Zuccotti M; SIERR. Maternal effect factors that contribute to oocytes developmental competence: an update. J Assist Reprod Genet. 2022 Apr;39(4):861-871. doi: 10.1007/s10815-022-02434-y. Epub 2022 Feb 15.

  PMID: 35165782 BACKGROUND

• Rienzi L, Vajta G, Ubaldi F. Predictive value of oocyte morphology in human IVF: a systematic review of the literature. Hum Reprod Update. 2011 Jan-Feb;17(1):34-45. doi: 10.1093/humupd/dmq029. Epub 2010 Jul 16.

  PMID: 20639518 BACKGROUND

• Tepla O, Topurko Z, Jirsova S, Moosova M, Fajmonova E, Cabela R, Komrskova K, Kratochvilova I, Masata J. Timing of ICSI with Respect to Meiotic Spindle Status. Int J Mol Sci. 2022 Dec 21;24(1):105. doi: 10.3390/ijms24010105.

  PMID: 36613547 RESULT

MeSH Terms

Conditions

Infertility; Infertility, Male; Infertility, Female

Interventions

Sperm Injections, Intracytoplasmic

Condition Hierarchy (Ancestors)

Genital Diseases; Urogenital Diseases; Genital Diseases, Male; Male Urogenital Diseases; Genital Diseases, Female; Female Urogenital Diseases; Female Urogenital Diseases and Pregnancy Complications

Intervention Hierarchy (Ancestors)

Fertilization in Vitro; Reproductive Techniques, Assisted; Reproductive Techniques; Therapeutics; Investigative Techniques

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: CARE PROVIDER
• Purpose: DIAGNOSTIC
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Leading doctor of the Center of Urogynecology and Pelvic Recontructive Surgery

Study Record Dates

• First Submitted: July 23, 2024
• First Posted: August 6, 2024
• Study Start: July 18, 2024
• Primary Completion (Estimated): December 31, 2027
• Study Completion (Estimated): March 31, 2028
• Last Updated: August 6, 2024

Record last verified: 2024-08

Locations

CZ (1)