Abstract
This study was to determine whether multiple vitrification-warming procedures and insemination method are associated with pregnancy and neonatal outcomes in preimplantation genetic testing for aneuploidy (PGT-A). This was a retrospective, single-center, observational study of 112 patients who underwent standard PGT-A practice and 154 patients who desired PGT-A for their vitrified unbiopsied blastocysts. A total of 97 euploid blastocysts biopsied and vitrified-warmed once and 117 euploid blastocysts biopsied once but vitrified-warmed twice (83 in vitro fertilization [IVF]–derived and 34 intracytoplasmic sperm injection [ICSI]–derived euploid blastocysts) were transferred. The primary outcome was the blastocyst survival rate for transfer, live birth rate, and neonatal outcomes. The results showed that an additional vitrification-warming procedure on blastocysts resulted in a lower but not statistically different survival rate for transfer. Compared with euploid blastocysts vitrified-warmed once, those vitrified-warmed twice provided statistically similar live birth rate. Neonatal outcomes, including the sex ratio, gestational age, birthweight, preterm birth rate, and low birthweight rate, did not differ between single and double vitrification. No significant differences were observed in rates of blastocyst survival, blastocyst euploid and live birth, and neonatal outcomes resulting from either conventional IVF or ICSI. The neonatal follow-up of babies live-born so far did not report any congenital malformations. In conclusion, an additional vitrification-warming on blastocysts had no detectable adverse impact on clinical outcomes after frozen-thawed single euploid blastocyst transfer in PGT-A cases; and ICSI did not confer any benefit in improving clinical outcomes compared with conventional IVF in cases requiring PGT-A on already vitrified nonbiopsied blastocysts.
Similar content being viewed by others
Data availability
Not applicable.
Code Availability
Not applicable.
References
Dahdouh EM. Preimplantation genetic testing for aneuploidy: a review of the evidence. Obstet Gynecol. 2021;137:528–34.
Neal SA, Morin SJ, Franasiak JM, Goodman LR, Juneau CR, Forman EJ, et al. Preimplantation genetic testing for aneuploidy is cost-effective, shortens treatment time, and reduces the risk of failed embryo transfer and clinical miscarriage. Fertil Steril. 2018;110:896–904.
Carvalho F, Coonen E, Goossens V, Kokkali G, Rubio C, Meijer-Hoogeveen M, et al. ESHRE PGT Consortium good practice recommendations for the organisation of PGT. Hum Reprod Open. 2020;2020:hoaa021.
Nagy ZP, Shapiro D, Chang CC. Vitrification of the human embryo: a more efficient and safer in vitro fertilization treatment. Fertil Steril. 2020;113:241–7.
Cimadomo D, Rienzi L, Romanelli V, Alviggi E, Levi-Setti PE, Albani E, et al. Inconclusive chromosomal assessment after blastocyst biopsy: prevalence, causative factors and outcomes after re-biopsy and re-vitrification. A multicenter experience. Hum Reprod. 2018;33:1839–46.
Tiegs AW, Tao X, Zhan Y, Whitehead C, Kim J, Hanson B, et al. A multicenter, prospective, blinded, nonselection study evaluating the predictive value of an aneuploid diagnosis using a targeted next-generation sequencing-based preimplantation genetic testing for aneuploidy assay and impact of biopsy. Fertil Steril. 2021;115:627–37.
He H, Jing S, Lu CF, Tan YQ, Luo KL, Zhang SP, et al. Neonatal outcomes of live births after blastocyst biopsy in preimplantation genetic testing cycles: a follow-up of 1,721 children. Fertil Steril. 2019;112:82–8.
Coates A, Kung A, Mounts E, Hesla J, Bankowski B, Barbieri E, et al. Optimal euploid embryo transfer strategy, fresh versus frozen, after preimplantation genetic screening with next generation sequencing: a randomized controlled trial. Fertil Steril. 2017;107:723-30.e3.
Harton GL, Magli MC, Lundin K, Montag M, Lemmen J, Harper JC. ESHRE PGD consortium/embryology special interest group–best practice guidelines for polar body and embryo biopsy for preimplantation genetic diagnosis/screening (PGD/PGS). Hum Reprod. 2011;26:41–6.
Practice Committees of the American Society for Reproductive Medicine and the Society for Assisted Reproductive Technology. Electronic address: asrm@asrm.org. Intracytoplasmic sperm injection (ICSI) for non-male factor indications: a committee opinion. Fertil Steril. 2020;114:239–45.
Palmerola KL, Vitez SF, Amrane S, Fischer CP, Forman EJ. Minimizing mosaicism: assessing the impact of fertilization method on rate of mosaicism after next-generation sequencing (NGS) preimplantation genetic testing for aneuploidy (PGT-A). J Assist Reprod Genet. 2019;36:153–7.
De Munck N, El Khatib I, Abdala A, El-Damen A, Bayram A, Arnanz A, et al. Intracytoplasmic sperm injection is not superior to conventional IVF in couples with non-male factor infertility and preimplantation genetic testing for aneuploidies (PGT-A). Hum Reprod. 2020;35:317–27.
Taylor TH, Patrick JL, Gitlin SA, Michael Wilson J, Crain JL, Griffin DK. Outcomes of blastocysts biopsied and vitrified once versus those cryopreserved twice for euploid blastocyst transfer. Reprod Biomed Online. 2014;29:59–64.
Bradley CK, Livingstone M, Traversa MV, McArthur SJ. Impact of multiple blastocyst biopsy and vitrification-warming procedures on pregnancy outcomes. Fertil Steril. 2017;108:999–1006.
De Vos A, Van Landuyt L, De Rycke M, Verdyck P, Verheyen G, Buysse A, et al. Multiple vitrification-warming and biopsy procedures on human embryos: clinical outcome and neonatal follow-up of children. Hum Reprod. 2020;35:2488–96.
Aluko A, Vaughan DA, Modest AM, Penzias AS, Hacker MR, Thornton K, et al. Multiple cryopreservation-warming cycles, coupled with blastocyst biopsy, negatively affect IVF outcomes. Reprod Biomed Online. 2021;42:572–8.
Shao Y, Li J, Lu J, Li H, Zhu Y, Jiang W, et al. Clinical outcomes of preimplantation genetic testing (PGT) application in couples with chromosomal inversion, a study in the Chinese Han population. Reprod Biol Endocrinol. 2020;18:79.
Gardner DK, Lane M, Stevens J, Schlenker T, Schoolcraft WB. Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertil Steril. 2000;73:1155–8.
Mazur P. Equilibrium, quasi-equilibrium, and nonequilibrium freezing of mammalian embryos. Cell Biophys. 1990;17:53–92.
Rienzi L, Gracia C, Maggiulli R, LaBarbera AR, Kaser DJ, Ubaldi FM, et al. Oocyte, embryo and blastocyst cryopreservation in ART: systematic review and meta-analysis comparing slow-freezing versus vitrification to produce evidence for the development of global guidance. Hum Reprod Update. 2017;23:139–55.
RezazadehValojerdi M, Eftekhari-Yazdi P, Karimian L, Hassani F, Movaghar B. Vitrification versus slow freezing gives excellent survival, post warming embryo morphology and pregnancy outcomes for human cleaved embryos. J Assist Reprod Genet. 2009;26:347–54.
Estudillo E, Jiménez A, Bustamante-Nieves PE, Palacios-Reyes C, Velasco I, López-Ornelas A. Cryopreservation of gametes and embryos and their molecular changes. Int J Mol Sci. 2021;22:10864.
Somoskoi B, Martino NA, Cardone RA, Lacalandra GM, Dell’Aquila ME, Cseh S. Different chromatin and energy/redox responses of mouse morulae and blastocysts to slow freezing and vitrification. Reprod Biol Endocrinol. 2015;13:22.
Li L, Zhang X, Zhao L, Xia X, Wang W. Comparison of DNA apoptosis in mouse and human blastocysts after vitrification and slow freezing. Mol Reprod Dev. 2012;79:229–36.
Carles M, Sonigo C, Binois O, Hesters L, Steffann J, Romana S, et al. Second biopsy for embryos with inconclusive results after preimplantation genetic testing: impact on pregnancy outcomes. J Gynecol Obstet Hum Reprod. 2022;51:102436.
Zacchini F, Arena R, Abramik A, Ptak GE. Embryo biopsy and development: the known and the unknown. Reproduction. 2017;154:R143–8.
Neal SA, Sun L, Jalas C, Morin SJ, Molinaro TA, Scott RT Jr. When next-generation sequencing-based preimplantation genetic testing for aneuploidy (PGT-A) yields an inconclusive report: diagnostic results and clinical outcomes after re biopsy. J Assist Reprod Genet. 2019;36:2103–9.
Ding M, Diao Z, Zhou J. The preimplantation genetic testing clinical outcomes of biopsy on vitrification-warming embryos: a retrospective study. J Obstet Gynaecol Res. 2022;48:1621–31.
Li X, Li W, Jia H, Gao Y, Shi W, Bai H. Double vitrification-warming cycles, coupled with blastocyst biopsy, impair live birth but do not affect neonatal outcomes. Int J Gynaecol Obstet. 2022. https://doi.org/10.1002/ijgo.14355.
Drakopoulos P, Garcia-Velasco J, Bosch E, Blockeel C, de Vos M, Santos-Ribeiro S, et al. ICSI does not offer any benefit over conventional IVF across different ovarian response categories in non-male factor infertility: a European multicenter analysis. J Assist Reprod Genet. 2019;36:2067–76.
Supramaniam PR, Granne I, Ohuma EO, Lim LN, McVeigh E, Venkatakrishnan R, et al. ICSI does not improve reproductive outcomes in autologous ovarian response cycles with non-male factor subfertility. Hum Reprod. 2020;35:583–94.
Liu L, Wang H, Li Z, Niu J, Tang R. Obstetric and perinatal outcomes of intracytoplasmic sperm injection versus conventional in vitro fertilization in couples with nonsevere male infertility. Fertil Steril. 2020;114:792–800.
Dang VQ, Vuong LN, Luu TM, Pham TD, Ho TM, Ha AN, et al. Intracytoplasmic sperm injection versus conventional in-vitro fertilisation in couples with infertility in whom the male partner has normal total sperm count and motility: an open-label, randomised controlled trial. Lancet. 2021;397:1554–63.
Acknowledgements
The authors expressed gratitude to all participants involved in this study.
Funding
This study was funded by the National Key Research and Development Program of China (2021YFC2700604), General Program of National Natural Science Foundation of China (82171648), and Youth Program of National Natural Science Foundation of China (81901556, 82101752).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics Approval
The study was approved by the Institutional Review Board of Center for Reproductive Medicine, Shandong University. Informed consent was obtained from all individual participants included in the study.
Consent to Participate
Not applicable.
Consent for Publication
All authors read and gave their consent for publication.
Competing Interests
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhang, Q., Yu, W., Jin, C. et al. Impact of Multiple Vitrification-Warming Procedures and Insemination Methods on Pregnancy and Neonatal Outcomes in Preimplantation Genetic Testing for Aneuploidy. Reprod. Sci. 30, 2302–2312 (2023). https://doi.org/10.1007/s43032-023-01177-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s43032-023-01177-0