Infertility; In Vitro Fertilization; Embryo Development
Conditions
Keywords
Assisted hatching, Day-3 vs Day-5, Blastocyst morphology, Trophectoderm biopsy, PGT-A, Aneuploidy, Mosaicism
Brief summary
This randomized controlled trial evaluates the effect of assisted hatching (AH) timing on embryo development and genetic outcomes in in vitro fertilization (IVF) cycles with preimplantation genetic testing for aneuploidy (PGT-A). Eligible patients undergoing IVF with PGT-A will have embryos randomized to receive assisted hatching either on Day-3 or Day-5 of culture, prior to trophectoderm biopsy. The primary outcome is the rate of aneuploid blastocysts. Secondary outcomes include blastocyst morphology, expansion stage, and the correlation between morphology and genetic results. The study aims to determine whether earlier or later assisted hatching affects embryo viability and the accuracy of genetic testing. The trial will provide evidence to optimize laboratory protocols in IVF centers.
Detailed description
This prospective randomized controlled trial compares two standardized laboratory procedures of assisted hatching (AH) timing in IVF cycles undergoing PGT-A. In Arm A, AH will be performed on Day-3 embryos; in Arm B, AH will be performed on Day-5 blastocysts prior to trophectoderm biopsy. A total of approximately 200 IVF cycles, involving around 650 embryos, will be included. Randomization will be conducted using block randomization. The primary outcome is the proportion of aneuploid blastocysts identified by next-generation sequencing (NGS). Secondary outcomes include blastocyst morphology (ICM, TE, expansion), biopsy efficiency, and correlation between morphological parameters and genetic outcomes. All procedures are part of the routine IVF-PGT workflow, and no additional risks to patients are introduced beyond standard care. The trial has been approved by the Institutional Review Board of 16A Ha Dong General Hospital.
Interventions
PROCEDUREAssisted Hatching
Laser-assisted zona drilling performed either on Day-3 or Day-5 of culture, depending on randomization arm.
Sponsors
16A Ha Dong General Hospital
Hoang Minh Ngan
Study design
Allocation
RANDOMIZED
Intervention model
PARALLEL
Primary purpose
TREATMENT
Masking
SINGLE (Outcomes Assessor)
Masking description
The PGT-A analysts evaluating the NGS results were blinded to the assisted hatching timing.
Intervention model description
Embryos randomized to receive assisted hatching on Day-3 (Arm A) or Day-5 (Arm B) prior to trophectoderm biopsy.
Eligibility
Sex/Gender
FEMALE
Age
20 Years to 45 Years
Healthy volunteers
No
Inclusion criteria
* IVF cycles with PGT-A * Oocyte age 20-45 years (donor cycles: donor's age) * AMH 1-10 ng/mL within 12 months * AFC ≥5 follicles (2-9 mm, both ovaries, day 2-5) * Controlled ovarian stimulation: antagonist or PPOS protocols * Stratified analysis by age (\<30, 30-34, 35-39, ≥40) * Sperm source: ejaculated samples prepared by density-gradient or swim-up * No surgical sperm retrieval * No advanced sperm selection (IMSI, PICSI, MACS)
Exclusion criteria
* History of ≥3 failed IVF cycles or ≥3 consecutive miscarriages * Indication for PGT-M or PGT-SR (monogenic disorders, translocations) * Severe medical/autoimmune conditions affecting oocytes/embryos (e.g. uncontrolled diabetes, active lupus) * No MII oocytes available for ICSI * Embryos not eligible for biopsy per SOP (early zona hatching, degeneration, poor expansion) * Sperm from surgical retrieval or advanced sperm selection (IMSI, PICSI, MACS) * PCOS diagnosed by Rotterdam criteria (≥2/3 features)
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Proportion of aneuploid blastocysts | At the time of trophectoderm biopsy and PGT-A result (Day 5-6 of embryo culture). | Percentage of blastocysts diagnosed as aneuploid by next-generation sequencing (NGS) after trophectoderm biopsy in each group (Day-3 vs Day-5 AH). |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Blastocyst morphology (ICM and TE grade) | Day 5-6 of culture | Comparison of blastocyst inner cell mass (ICM) and trophectoderm (TE) grading between Day-3 and Day-5 AH groups. |
| Mosaicism rate | Day 5-6 of culture | Proportion of blastocysts diagnosed as mosaic by NGS in each group. |
Other
| Measure | Time frame | Description |
|---|---|---|
| Correlation between blastocyst morphology and genetic outcome | Day 5-6 of culture (at the time of trophectoderm biopsy and PGT-A result) | Analysis of the correlation between blastocyst morphology (ICM and TE grade, expansion stage) and genetic outcomes (euploid, aneuploid, mosaic). |
Countries
Vietnam
Contacts
Primary ContactHOANG M NGAN, MSc
Outcome results
None listed