Multiple cryopreservation-warming cycles, coupled with blastocyst biopsy, negatively affect IVF outcomes
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Updated: Nov 19, 2025

Human Blastocyst Biopsy and Vitrification
Published on: July 26, 2019
Ashley Aluko1, Denis A Vaughan2, Anna M Modest1
1Department of Obstetrics and Gynecology, Beth Israel Deaconess Medical Center, Boston MA, USA; Department of Obstetrics, Gynecology, and Reproductive Biology, Harvard Medical School, Boston MA, USA.
Research Question:
Do multiple cryopreservation-warming cycles, coupled with blastocyst biopsy, negatively affect IVF outcomes?
Design:
Patients undergoing IVF with homologous single embryo transfer, and who underwent trophectoderm biopsy for preimplantation genetic testing for aneuploidy (PGT-A) between 2013 and 2017, were divided into three groups based on degree of embryonic micromanipulation: once-biopsied, once-cryopreserved (group BC, n = 2603), once-biopsied, twice-cryopreserved (group CBC, n = 95) and twice-biopsied, twice-cryopreserved (group BCBC, n = 15). The primary outcome was live birth; secondary outcomes included positive serum pregnancy test, clinical pregnancy and miscarriage.
Results:
Group CBC had a significantly lower chance of live birth (adjusted RR 0.57, 95% CI 0.41 to 0.79) and clinical pregnancy (adjusted RR 0.67, 95% CI 0.53 to 0.85) compared with group BC. Miscarriage rates were similar between groups BC and CBC (adjusted RR 1.3, 95% CI 0.64 to 2.7).
Conclusions:
Multiple cryopreservation-warming cycles, coupled with blastocyst biopsy, negatively affect IVF outcomes. Although PGT-A is thought to improve reproductive outcomes on a per transfer basis, caution must be exercised in counselling patients on the possibility of diminishing returns owing to further embryonic micromanipulation after an embryo has been cryopreserved.
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