Infertility; In Vitro Fertilization; Embryo DevelopmentAH-PGTANCT07154875
Day-3 vs Day-5 Assisted Hatching: Impact on Blastocyst Morphology and PGT-A Outcomes
A study testing Laser-Assisted Hatching in people with Infertility; In Vitro Fertilization; Embryo Development, run by Hoang Minh Ngan at 1 site in Vietnam.
Trial summary
This randomized controlled trial compared assisted hatching (AH) performed on Day 3 versus Day 5 in patients undergoing IVF-ICSI with preimplantation genetic testing for aneuploidy (PGT-A). Read more about this study
- Who is it for?
- Ages 18 years and older, women only
- Conditions the study is for
- Infertility; In Vitro Fertilization; Embryo Development
- What is being tested?
- Laser-Assisted Hatching
- Type of treatment
- Surgery or procedure
- Could I receive a placebo?
- No, everyone receives an active treatment
- How many people
- 141 participants
- Where
- 1 siteVietnam
- Study dates
- 14 Jun 2025to 7 Jul 2026
- Phase
- Interventional
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 years and older
- Women only
- No healthy volunteers
- Adult, Older adult
Who may be able to join
- Female participants undergoing IVF-ICSI with an indication for PGT-A
- Age ≥18 years; no prespecified upper age limit. Age was recorded for subsequent analyses
- Anti-Müllerian hormone (AMH) level ≥1.0 ng/mL
- Antral follicle count (AFC) ≥5 follicles
- Controlled ovarian stimulation using an antagonist or progestin-primed ovarian stimulation (PPOS) protocol
- Use of ejaculated sperm prepared by density-gradient centrifugation or swim-up
- Written informed consent for participation
Who cannot join
- History of ≥3 failed IVF cycles or ≥3 consecutive miscarriages
- Indication for PGT-M or PGT-SR, including monogenic disorders or structural chromosomal rearrangements
- Severe uncontrolled medical or autoimmune conditions considered likely to affect oocyte or embryo development
- No metaphase II (MII) oocytes available for ICSI
- Use of surgically retrieved sperm
- Use of advanced sperm-selection techniques, including IMSI, PICSI, or MACS
- Polycystic ovary syndrome diagnosed according to the Rotterdam criteria
Trial locations and contacts
1 site in 1 country.
Vietnam1 site
Center for Reproductive Support, 16A Ha Dong General Hospital
Hanoi, Hanoi, 100000
About this study
Official title: Randomized Controlled Trial of Assisted Hatching Timing (Day-3 vs Day-5) and Its Impact on Blastocyst Morphology and Genetic Outcomes in IVF Cycles Undergoing PGT-A
This randomized controlled trial compared assisted hatching (AH) performed on Day 3 versus Day 5 in patients undergoing IVF-ICSI with preimplantation genetic testing for aneuploidy (PGT-A). Eligible IVF-ICSI cycles were randomly assigned to one of the two AH strategies, and all embryos from each randomized cycle followed the assigned strategy. The study evaluated whether the timing of AH was associated with differences in blastocyst development, blastocyst morphology, and PGT-A outcomes. The findings are intended to help optimize the timing of assisted hatching in IVF laboratory practice.
Read the detailed description
This prospective randomized controlled trial compared two standardized assisted-hatching strategies in IVF-ICSI cycles undergoing preimplantation genetic testing for aneuploidy (PGT-A). Eligible IVF-ICSI cycles were randomly allocated in a 1:1 ratio to Day-3 assisted hatching (Arm A) or Day-5 assisted hatching (Arm B). Randomization was performed at the IVF-ICSI cycle level using fixed block randomization with a block size of 4. Randomization was not stratified by maternal age, anti-Müllerian hormone level, or other clinical characteristics. Women who underwent more than one eligible IVF-ICSI cycle could contribute more than one randomized cycle, with each eligible cycle assigned according to the randomization sequence. All embryos derived from each randomized cycle followed the assigned assisted-hatching strategy.
In Arm A, laser-assisted hatching was performed on Day-3 embryos. In Arm B, laser-assisted hatching was performed on Day-5 blastocysts. In both groups, blastocysts were evaluated on Day 5 and subsequently cultured for approximately 6-7 additional hours before trophectoderm biopsy. This interval was standardized across both study arms and was consistent with the routine laboratory workflow, while allowing additional time for blastocyst expansion and hatching before biopsy. Blastocyst morphology used for the primary morphological analyses was assessed at the time of trophectoderm biopsy according to standardized criteria, including blastocyst expansion stage, inner cell mass (ICM) morphology, and trophectoderm (TE) morphology.
Blastocysts meeting biopsy criteria underwent trophectoderm biopsy followed by PGT-A using next-generation sequencing. PGT-A was performed using paired-end sequencing with the EmbryoMap™ Kit (Vitrolife) on the Illumina MiSeq sequencing system. PGT-A testing and interpretation were conducted by a separate genetics laboratory whose analysts were blinded to the assisted-hatching allocation.
A total of 141 women contributed 158 randomized IVF-ICSI cycles to the study, with 79 cycles allocated to Day-3 assisted hatching and 79 cycles allocated to Day-5 assisted hatching. Sixteen women contributed more than one IVF-ICSI cycle: 15 women contributed two cycles and one woman contributed three cycles. Statistical analyses accounted for within-participant correlation arising from repeated IVF-ICSI cycles and multiple embryos contributed by the same woman, where appropriate.
Participants and treating clinicians were unaware of the assigned assisted-hatching timing. The embryologist performing assisted hatching could not be blinded at the time of the intervention because knowledge of the assigned timing was required to perform the procedure.
- Being tested
- Laser-Assisted Hatching
- Keywords
- Assisted hatching, Day-3 vs Day-5, Blastocyst morphology, Trophectoderm biopsy, PGT-A, Aneuploidy, Mosaicism
- Medical subject headings
- Infertility, Aneuploidy
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Laser-Assisted HatchingProcedure or surgery
Also called Assisted Hatching (AH), Laser-Assisted Hatching (LAH)
A standardized laser-assisted opening of the zona pellucida was performed either on Day 3 or Day 5 according to randomized group assignment. The only protocol-defined difference between study arms was the timing of assisted hatching; subsequent blastocyst culture, Day-5 assessment, and trophectoderm biopsy procedures were standardized across both groups.
| Study group | Role | What they receive |
|---|---|---|
| Day-3 Assisted Hatching | Experimental | Laser-assisted hatching was performed on Day-3 embryos according to the standardized laboratory protocol. Embryos were then cultured to the blastocyst stage. On Day 5, blastocysts were evaluated and cultured for approximately 6-7 additional hours before trophectoderm biopsy for PGT-A. Receives: Procedure: Laser-Assisted Hatching |
| Day-5 Assisted Hatching | Experimental | Embryos were cultured without assisted hatching until Day 5. Laser-assisted hatching was then performed on Day-5 blastocysts according to the standardized laboratory protocol. Blastocysts were subsequently cultured for approximately 6-7 additional hours before trophectoderm biopsy for PGT-A. Receives: Procedure: Laser-Assisted Hatching |
What is being measured
The primary outcome is the main question the study is designed to answer.
Distribution of PGT-A Genetic Classifications
Blastocysts with a valid PGT-A result were classified into three mutually exclusive categories: euploid, mosaic, or abnormal. The distribution of these three PGT-A classifications was compared between the Day-3 and Day-5 assisted-hatching groups.
Measured: Following PGT-A of blastocysts biopsied on Day 5 or Day 6 of embryo culture
Blastocyst Morphology at Trophectoderm Biopsy
Blastocyst morphology was assessed at the time of trophectoderm biopsy using standardized criteria, including blastocyst expansion stage, inner cell mass (ICM) morphology, and trophectoderm (TE) morphology. The distributions of these morphological characteristics were compared between the Day-3 and Day-5 assisted-hatching groups.
Measured: At trophectoderm biopsy on Day 5 or Day 6 of embryo culture
Secondary outcomes (1)
Proportion of Mosaic Blastocysts
Blastocysts with a valid PGT-A result were classified as euploid, mosaic, or abnormal. The proportion of blastocysts classified as mosaic was calculated for each assisted-hatching group, using the number of blastocysts with a valid PGT-A result as the denominator.
Measured: Following PGT-A of blastocysts biopsied on Day 5 or Day 6 of embryo culture
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted4 Sep 2025
- Study start14 Jun 2025
- Primary completion7 Jul 2026
- Study completion7 Jul 2026
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (3)Papers the sponsor has linked to this study.
- Other trials for Infertility; In Vitro Fertilization; Embryo DevelopmentStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Treatment
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Model description
- Eligible participants undergoing IVF-ICSI were randomly allocated in a 1:1 ratio to Day-3 assisted hatching (Arm A) or Day-5 assisted hatching (Arm B). Randomization was performed at the participant/IVF-ICSI cycle level, and all embryos derived from each randomized cycle followed the assigned assisted-hatching strategy.
- Masking
- Triple blind
- Who is masked
- Participant, Care provider, Outcomes assessor
- Enrolment
- 141 participants (actual)
Sponsor and investigators
- Lead sponsor
- Hoang Minh Ngan
- Sponsor type
- University, hospital or other
- Collaborators
- 16A Ha Dong General Hospital
- Responsible party
- Sponsor-investigator, Hoang Minh Ngan, 16A Ha Dong General Hospital
- Investigators
- Ngan M Hoang, MSc, Study chair, Center for Reproductive Support, 16A Ha Dong General HospitalTao D Nguyen, MD, PhD, Professor, Principal investigator, Center for Reproductive Support, 16A Ha Dong General HospitalTon D Nguyen, PhD, Principal investigator, Institute of Biology, Vietnam Academy of Science and TechnologySon T Trinh, MD, PhD, Principal investigator, Vietnam Military Medical University, Ministry of National Defence
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07154875
- Sponsor's protocol ID
- AH-PGTA-2025
- Other IDs
- 16A-IRB-2025
- Acronym
- AH-PGTA
- Status last verified
- Sep 2026
- First submitted
- 27 Aug 2025
- Last updated
- 10 Sep 2026
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No: The plan for sharing individual participant data (IPD) has not yet been finalized. De-identified participant- and embryo-level data may be considered for sharing upon reasonable request for academic research, subject to applicable ethical, institutional, and data-protection requirements.
Publications
ESHRE Working Group on Recurrent Implantation Failure; Cimadomo D, de Los Santos MJ, Griesinger G, Lainas G, Le Clef N, McLernon DJ, Montjean D, Toth B, Vermeulen N, Macklon N. ESHRE good practice recommendations on recurrent implantation failure. Hum Reprod Open. 2023 Jun 15;2023(3):hoad023. doi: 10.1093/hropen/hoad023. eCollection 2023.
PubMed 37332387McArthur SJ, Leigh D, Marshall JT, de Boer KA, Jansen RP. Pregnancies and live births after trophectoderm biopsy and preimplantation genetic testing of human blastocysts. Fertil Steril. 2005 Dec;84(6):1628-36. doi: 10.1016/j.fertnstert.2005.05.063.
PubMed 16359956Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, Nagy ZP, Ubaldi FM. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014 Jun;29(6):1173-81. doi: 10.1093/humrep/deu033. Epub 2014 Feb 26.
PubMed 24578475
Study at a glance
From the registry record, last updated by the sponsor on 10 Sep 2026. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Randomized Controlled Trial of Assisted Hatching Timing (Day-3 vs Day-5) and Its Impact on Blastocyst Morphology and Genetic Outcomes in IVF Cycles Undergoing PGT-A
- ClinicalTrials.gov ID
- NCT07154875
- Acronym / sponsor ID
- AH-PGTA / AH-PGTA-2025
- Keywords
- Assisted hatching, Day-3 vs Day-5, Blastocyst morphology, Trophectoderm biopsy, PGT-A, Aneuploidy, Mosaicism
- Study type
- Interventional
- Recruitment status
- Completed
- Enrolment
- 141 participants (actual)
- Lead sponsor
- Hoang Minh NganUniversity, hospital or other
- Collaborators
- 16A Ha Dong General Hospital
- Study start
- 14 Jun 2025 (actual)
- Primary completion
- 7 Jul 2026 (actual)
- Study completion
- 7 Jul 2026 (actual)
- First posted
- 4 Sep 2025 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
This randomized controlled trial compared assisted hatching (AH) performed on Day 3 versus Day 5 in patients undergoing IVF-ICSI with preimplantation genetic testing for aneuploidy (PGT-A). Eligible IVF-ICSI cycles were randomly assigned to one of the two AH strategies, and all embryos from each randomized cycle followed the assigned strategy. The study evaluated whether the timing of AH was associated with differences in blastocyst development, blastocyst morphology, and PGT-A outcomes. The findings are intended to help optimize the timing of assisted hatching in IVF laboratory practice.
Detailed description
This prospective randomized controlled trial compared two standardized assisted-hatching strategies in IVF-ICSI cycles undergoing preimplantation genetic testing for aneuploidy (PGT-A). Eligible IVF-ICSI cycles were randomly allocated in a 1:1 ratio to Day-3 assisted hatching (Arm A) or Day-5 assisted hatching (Arm B). Randomization was performed at the IVF-ICSI cycle level using fixed block randomization with a block size of 4. Randomization was not stratified by maternal age, anti-Müllerian hormone level, or other clinical characteristics. Women who underwent more than one eligible IVF-ICSI cycle could contribute more than one randomized cycle, with each eligible cycle assigned according to the randomization sequence. All embryos derived from each randomized cycle followed the assigned assisted-hatching strategy.
In Arm A, laser-assisted hatching was performed on Day-3 embryos. In Arm B, laser-assisted hatching was performed on Day-5 blastocysts. In both groups, blastocysts were evaluated on Day 5 and subsequently cultured for approximately 6-7 additional hours before trophectoderm biopsy. This interval was standardized across both study arms and was consistent with the routine laboratory workflow, while allowing additional time for blastocyst expansion and hatching before biopsy. Blastocyst morphology used for the primary morphological analyses was assessed at the time of trophectoderm biopsy according to standardized criteria, including blastocyst expansion stage, inner cell mass (ICM) morphology, and trophectoderm (TE) morphology.
Blastocysts meeting biopsy criteria underwent trophectoderm biopsy followed by PGT-A using next-generation sequencing. PGT-A was performed using paired-end sequencing with the EmbryoMap™ Kit (Vitrolife) on the Illumina MiSeq sequencing system. PGT-A testing and interpretation were conducted by a separate genetics laboratory whose analysts were blinded to the assisted-hatching allocation.
A total of 141 women contributed 158 randomized IVF-ICSI cycles to the study, with 79 cycles allocated to Day-3 assisted hatching and 79 cycles allocated to Day-5 assisted hatching. Sixteen women contributed more than one IVF-ICSI cycle: 15 women contributed two cycles and one woman contributed three cycles. Statistical analyses accounted for within-participant correlation arising from repeated IVF-ICSI cycles and multiple embryos contributed by the same woman, where appropriate.
Participants and treating clinicians were unaware of the assigned assisted-hatching timing. The embryologist performing assisted hatching could not be blinded at the time of the intervention because knowledge of the assigned timing was required to perform the procedure.
Study design
- Primary purpose
- Treatment
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Masking
- Triple blind (participant, care provider, outcomes assessor)
- Number of arms
- 2
- Model description
- Eligible participants undergoing IVF-ICSI were randomly allocated in a 1:1 ratio to Day-3 assisted hatching (Arm A) or Day-5 assisted hatching (Arm B). Randomization was performed at the participant/IVF-ICSI cycle level, and all embryos derived from each randomized cycle followed the assigned assisted-hatching strategy.
| Arm | Type | Receives |
|---|---|---|
| Day-3 Assisted Hatching | Experimental | Procedure: Laser-Assisted HatchingLaser-assisted hatching was performed on Day-3 embryos according to the standardized laboratory protocol. Embryos were then cultured to the blastocyst stage. On Day 5, blastocysts were evaluated and cultured for approximately 6-7 additional hours before trophectoderm biopsy for PGT-A. |
| Day-5 Assisted Hatching | Experimental | Procedure: Laser-Assisted HatchingEmbryos were cultured without assisted hatching until Day 5. Laser-assisted hatching was then performed on Day-5 blastocysts according to the standardized laboratory protocol. Blastocysts were subsequently cultured for approximately 6-7 additional hours before trophectoderm biopsy for PGT-A. |
Who may qualify
- Ages
- 18 years and older
- Sex
- Female only
- Healthy volunteers
- Not accepted
- Age groups
- Adult, Older adult
Key inclusion criteria (7 listed)
- Female participants undergoing IVF-ICSI with an indication for PGT-A
- Age ≥18 years; no prespecified upper age limit. Age was recorded for subsequent analyses
- Anti-Müllerian hormone (AMH) level ≥1.0 ng/mL
- Antral follicle count (AFC) ≥5 follicles
- Controlled ovarian stimulation using an antagonist or progestin-primed ovarian stimulation (PPOS) protocol
Key exclusion criteria (7 listed)
- History of ≥3 failed IVF cycles or ≥3 consecutive miscarriages
- Indication for PGT-M or PGT-SR, including monogenic disorders or structural chromosomal rearrangements
- Severe uncontrolled medical or autoimmune conditions considered likely to affect oocyte or embryo development
- No metaphase II (MII) oocytes available for ICSI
- Use of surgically retrieved sperm
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Laser-Assisted HatchingAlso: Assisted Hatching (AH), Laser-Assisted Hatching (LAH)A standardized laser-assisted opening of the zona pellucida was performed either on Day 3 or Day 5 according to randomized group assignment. The only protocol-defined difference between study arms was the timing of assisted hatching; subsequent blastocyst culture, Day-5 assessment, and trophectoderm biopsy procedures were standardized across both groups. | Procedure or surgery | Day-3 Assisted Hatching, Day-5 Assisted Hatching |
Primary outcome
Distribution of PGT-A Genetic Classifications
Time frame: Following PGT-A of blastocysts biopsied on Day 5 or Day 6 of embryo culture
Blastocysts with a valid PGT-A result were classified into three mutually exclusive categories: euploid, mosaic, or abnormal. The distribution of these three PGT-A classifications was compared between the Day-3 and Day-5 assisted-hatching groups.
Blastocyst Morphology at Trophectoderm Biopsy
Time frame: At trophectoderm biopsy on Day 5 or Day 6 of embryo culture
Blastocyst morphology was assessed at the time of trophectoderm biopsy using standardized criteria, including blastocyst expansion stage, inner cell mass (ICM) morphology, and trophectoderm (TE) morphology. The distributions of these morphological characteristics were compared between the Day-3 and Day-5 assisted-hatching groups.
1 secondary outcome
Proportion of Mosaic Blastocysts
Time frame: Following PGT-A of blastocysts biopsied on Day 5 or Day 6 of embryo culture
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 10 Sep 2026.
- Background: PubMed 37332387ESHRE Working Group on Recurrent Implantation Failure; Cimadomo D, de Los Santos MJ, Griesinger G, Lainas G, Le Clef N, McLernon DJ, Montjean D, Toth B, Vermeulen N, Macklon N. ESHRE good practice recommendations on recurrent implantation failure. Hum Reprod Open. 2023 Jun 15;2023(3):hoad023. doi: 10.1093/hropen/hoad023. eCollection 2023.
- Background: PubMed 16359956McArthur SJ, Leigh D, Marshall JT, de Boer KA, Jansen RP. Pregnancies and live births after trophectoderm biopsy and preimplantation genetic testing of human blastocysts. Fertil Steril. 2005 Dec;84(6):1628-36. doi: 10.1016/j.fertnstert.2005.05.063.
- Background: PubMed 24578475Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, Nagy ZP, Ubaldi FM. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014 Jun;29(6):1173-81. doi: 10.1093/humrep/deu033. Epub 2014 Feb 26.
- Official website of 16A Ha Dong General Hospital - Center for Reproductive SupportLink provided by the sponsor.
- Individual participant dataSharing planned: No. The plan for sharing individual participant data (IPD) has not yet been finalized. De-identified participant- and embryo-level data may be considered for sharing upon reasonable request for academic research, subject to applicable ethical, institutional, and data-protection requirements.
Related trials for Infertility; In Vitro Fertilization; Embryo Development
Recruiting
Day-4 Embryo Defragmentation: Blastocyst Rate and clInical Outcomes
Female Infertility, Embryo Development
Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Ngan M Hoang, MSc study chair
Center for Reproductive Support, 16A Ha Dong General Hospital
Tao D Nguyen, MD, PhD, Professor principal investigator
Center for Reproductive Support, 16A Ha Dong General Hospital
Ton D Nguyen, PhD principal investigator
Institute of Biology, Vietnam Academy of Science and Technology
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
A study testing Laser-Assisted Hatching in people with Infertility; In Vitro Fertilization; Embryo Development, run by Hoang Minh Ngan at 1 site in Vietnam.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 7 inclusion and 7 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 10 Sep 2026. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Vietnam. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Distribution of PGT-A Genetic Classifications (time frame: Following PGT-A of blastocysts biopsied on Day 5 or Day 6 of embryo culture). The study lists 2 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT07154875, which also lists 3 linked publications.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 10 Sep 2026; AzTrials last refreshed its copy on 30 Oct 2026.