Day-3 vs Day-5 Assisted Hatching: Impact on Blastocyst Morphology and PGT-A Outcomes (AH-PGTA)

Hoang Minh Ngan

Status: Enrolling

Conditions: Infertility; In Vitro Fertilization; Embryo Development

Treatments: Procedure: Assisted Hatching

Study type: Interventional

Funder types: Other

Identifiers

NCT07154875

16A-IRB-2025 (Other Identifier)

AH-PGTA-2025

Details and patient eligibility

About

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.

Full 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.

Enrollment: 200 estimated patients

Sex: Female

Ages: 20 to 45 years old

Volunteers: No Healthy Volunteers

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)

Trial design

Primary purpose: Treatment

Allocation: Randomized

Interventional model: Parallel Assignment

Masking: Single Blind

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200 participants in 2 patient groups

Day-3 Assisted Hatching

Experimental group

Description:

Embryos undergo laser-assisted zona drilling on Day-3. Embryos are then cultured to Day-5 and biopsied for PGT-A.

Treatment:

Procedure: Assisted Hatching

Day-5 Assisted Hatching

Experimental group

Description:

Embryos are cultured without zona drilling until Day-5. Laser-assisted hatching is performed immediately before trophectoderm biopsy for PGT-A.

Treatment:

Procedure: Assisted Hatching