Embryonic and Neonatal Outcomes Following Double Vitrification/thawing: a Systematic Review and Meta-analysis

Overview

Journal BMC Pregnancy Childbirth

Publisher Biomed Central

Specialty Gynecology & Obstetrics

Date 2025 Feb 26

PMID 40012071

Authors

Arezoo Maleki-Hajiagha

Anahid Shafie

Saeede Rezayi

Mahnaz Marvi

Rana Karimi

Fardin Amidi

Affiliations

Soon will be listed here.

Abstract

Objectives: This systematic review and meta-analysis aimed to evaluate the impact of double vitrification/thawing (DVT) versus single vitrification/thawing (SVT) on key embryonic and neonatal outcomes.

Data Extraction: Information sources included systematic search in PubMed, Scopus, and Cochrane databases up to September 7, 2024. Data from each qualifying study were extracted by two reviewers using a standardized electronic data gathering form.

Data Analysis: Mantel-Haenszel odds ratio (MHOR) and mean difference (MD) with 95% confidence intervals (CI) were calculated using both fixed and random-effects models. Subgroup analyses were based on biopsy status, number of biopsy rounds, extended culture between rounds of vitrification, and embryo transfer strategy.

Results: A total of 35 studies involving 46,749 embryo transfer cycles were included. After excluding studies that used slow freezing, 28 studies were included in the meta-analyses. The findings indicated that DVT is associated with significant reductions in cryosurvival rates (MHOR: 0.4; CI: 0.3 to 0.8; P < 0.01), biochemical pregnancy (MHOR: 0.7; CI: 0.6 to 0.8; P < 0.01), clinical pregnancy (MHOR: 0.7; CI: 0.5 to 0.8; P < 0.01), and live birth rates (MHOR: 0.6; CI: 0.5 to 0.7; P < 0.01). Additionally, there was a significant increase in the miscarriage rate (MHOR: 1.4; CI: 1.2 to 1.7; P < 0.01). No significant differences were found in neonatal outcomes.

Conclusion: Poor-quality evidence suggests that the transfer of double-vitrified embryos might be associated with significantly lower rates of cryosurvival, pregnancy, and live births; however, it does not appear to affect neonatal outcomes such as birth weight and gestational age at birth. Given the small sample size in some subgroups, the high risk of selection, confounding and missing data biases, and the high level of heterogeneity for some outcomes, these findings should be interpreted cautiously.

References

Hernandez-Nieto C, Lee J, Alkon-Meadows T, Briton-Jones C, Sandler B, Copperman A . Biological relevance of trophectoderm morphology: initial β-hCG measurements correlate with trophectoderm grading on euploid frozen embryo transfers. J Assist Reprod Genet. 2022; 39(9):2051-2059. PMC: 9475011. DOI: 10.1007/s10815-022-02553-6. View

Carvalho F, Coonen E, Goossens V, Kokkali G, Rubio C, Meijer-Hoogeveen M . ESHRE PGT Consortium good practice recommendations for the organisation of PGT. Hum Reprod Open. 2020; 2020(3):hoaa021. PMC: 7257038. DOI: 10.1093/hropen/hoaa021. View

Page M, McKenzie J, Bossuyt P, Boutron I, Hoffmann T, Mulrow C . The PRISMA 2020 statement: an updated guideline for reporting systematic reviews. BMJ. 2021; 372:n71. PMC: 8005924. DOI: 10.1136/bmj.n71. View

Li Piani L, Petrone P, Brutto M, De Vos A, Van Der Kelen A, Vaiarelli A . A systematic review and meta-analysis of double trophectoderm biopsy and/or cryopreservation in PGT: balancing the need for a diagnosis against the risk of harm. Hum Reprod Update. 2024; 31(2):102-115. DOI: 10.1093/humupd/dmae031. View

Zolfaroli I, Romeu Villarroya M, Serralta Garcia L, Rubio Rubio J, Monzo Miralles A . Impact of prolonged embryo storage on reproductive and neonatal outcomes: a systematic review and meta-analysis. J Assist Reprod Genet. 2024; 41(10):2691-2700. PMC: 11534939. DOI: 10.1007/s10815-024-03283-7. View

Wei D, Legro R, Chen Z . The cumulative live birth rate after a freeze-only strategy versus a conventional fresh embryo transfer strategy: a call for more level 1 evidence. BMC Med. 2020; 18(1):12. PMC: 6977243. DOI: 10.1186/s12916-019-1479-2. View

Li J, Xiong S, Zhao Y, Li C, Han W, Huang G . Effect of the Re-Vitrification of Embryos at Different Stages on Embryonic Developmental Potential. Front Endocrinol (Lausanne). 2021; 12:653310. PMC: 8317612. DOI: 10.3389/fendo.2021.653310. View

. Indications and management of preimplantation genetic testing for monogenic conditions: a committee opinion. Fertil Steril. 2023; 120(1):61-71. DOI: 10.1016/j.fertnstert.2023.03.003. View

Stanger J, Wong J, Conceicao J, Yovich J . Vitrification of human embryos previously cryostored by either slow freezing or vitrification results in high pregnancy rates. Reprod Biomed Online. 2012; 24(3):314-20. DOI: 10.1016/j.rbmo.2011.11.013. View

10.

Oraiopoulou C, Karagianni M, Papatheodorou A, Toumpa O, Papadopoulou M, Christophoridis N . Double vitrification of embryos adversely affects clinical outcomes. JBRA Assist Reprod. 2024; 28(3):399-404. PMC: 11349271. DOI: 10.5935/1518-0557.20240014. View

11.

Miwa A, Noguchi Y, Hosoya K, Mori Y, Sato T, Kasahara Y . Equivalent clinical outcome after vitrified-thawed blastocyst transfer using semi-automated embryo vitrification system compared with manual vitrification method. Reprod Med Biol. 2020; 19(2):164-170. PMC: 7138946. DOI: 10.1002/rmb2.12320. View

12.

Theodorou E, Chronopoulou E, Ozturk O, Brunetti X, Serhal P, Ben-Nagi J . Impact of double trophectoderm biopsy on reproductive outcomes following single euploid blastocyst transfer. Eur J Obstet Gynecol Reprod Biol. 2024; 298:35-40. DOI: 10.1016/j.ejogrb.2024.04.033. View

13.

Farhi J, Elizur S, Yonish M, Seidman D, Shulman A, Schiff E . Assessment of a double freezing approach in the management of surplus embryos in IVF. Reprod Biomed Online. 2019; 38(4):517-519. DOI: 10.1016/j.rbmo.2018.11.010. View

14.

Barberet J, Ducreux B, Bruno C, Guilleman M, Simonot R, Lieury N . Comparison of oocyte vitrification using a semi-automated or a manual closed system in human siblings: survival and transcriptomic analyses. J Ovarian Res. 2022; 15(1):128. PMC: 9720994. DOI: 10.1186/s13048-022-01064-3. View

15.

Neal S, Sun L, Jalas C, Morin S, Molinaro T, Scott Jr R . 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(10):2103-2109. PMC: 6823328. DOI: 10.1007/s10815-019-01550-6. View

16.

Kopeika J, Thornhill A, Khalaf Y . The effect of cryopreservation on the genome of gametes and embryos: principles of cryobiology and critical appraisal of the evidence. Hum Reprod Update. 2014; 21(2):209-27. DOI: 10.1093/humupd/dmu063. View

17.

Check J, Brittingham D, Swenson K, Wilson C, Lurie D . Transfer of refrozen twice-thawed embryos do not decrease the implantation rate. Clin Exp Obstet Gynecol. 2001; 28(1):14-6. View

18.

Zhou G, Zeng Y, Guo J, Meng Q, Meng Q, Jia G . Vitrification transiently alters Oct-4, Bcl2 and P53 expression in mouse morulae but does not affect embryo development in vitro. Cryobiology. 2016; 73(2):120-5. DOI: 10.1016/j.cryobiol.2016.08.011. View

19.

Taylor T, Patrick J, Gitlin S, Wilson J, Crain J, Griffin D . Outcomes of blastocysts biopsied and vitrified once versus those cryopreserved twice for euploid blastocyst transfer. Reprod Biomed Online. 2014; 29(1):59-64. DOI: 10.1016/j.rbmo.2014.03.001. View

20.

Khatun H, Ihara Y, Takakura K, Egashira J, Wada Y, Konno T . Role of endoplasmic reticulum stress on developmental competency and cryo-tolerance in bovine embryos. Theriogenology. 2019; 142:131-137. DOI: 10.1016/j.theriogenology.2019.09.042. View