Abstract
Purpose
The purpose of this study is to investigate the application value of the extended embryo culture for 7–8 h in day 3 morning during IVF-ET process.
Methods
Embryos were retrospectively assessed during 08:00–09:00 on the morning of day 3 in the control group, and were assessed once again at 16:00 in the afternoon in the extended culture (EC) group. The embryos with good developmental potential were preferentially selected to transfer. The cumulative pregnancy outcomes were analyzed in one oocyte retrieval cycle.
Results
Similar proportions were found in the rates of cumulative clinical pregnancy, cumulative live birth, and the perinatal/neonatal outcomes per oocyte retrieval cycle (P > 0.05). But higher total clinical pregnancy rate, higher total implantation rate, and lower total abortion rate were obtained in the EC group (P < 0.05). After EC, 53.58% of the embryos were able to continue to develop. The transferred embryos were mainly composed of ≥ 8-cell embryos (75.90%) in the EC group and ≤ 8-cell embryos (82.92%) in the control group. Interestingly, the implantation rates were increasingly improved with the increasing blastomere number up to 56.31% at the morula stage in the EC group, while they were limited to 32.33% at 8-cell stage in the control group.
Conclusions
The extended culture of day 3 embryos for 7–8 h not only reduced the risk of IVF-ET treatment compared to blastocyst culture through another 2–3 days, but also improved the clinical outcomes and the efficiency of every transferred cycle and every transferred embryo.
Similar content being viewed by others
References
Braude P, Bolton V, Moore S. Human gene expression first occurs between the four- and eight-cell stages of preimplantation development. Nature. 1988;332(6163):459–61. https://doi.org/10.1038/332459a0.
Assou S, Boumela I, Haouzi D, Anahory T, Dechaud H, De Vos J, et al. Dynamic changes in gene expression during human early embryo development: from fundamental aspects to clinical applications. Hum Reprod Update. 2011;17(2):272–90. https://doi.org/10.1093/humupd/dmq036.
Paternot G, Wetzels AM, Thonon F, Vansteenbrugge A, Willemen D, Devroe J, et al. Intra- and interobserver analysis in the morphological assessment of early stage embryos during an IVF procedure: a multicentre study. Reprod Biol Endocrinol. 2011;9:127. https://doi.org/10.1186/1477-7827-9-127.
Shi W, Xue X, Zhang S, Zhao W, Liu S, Zhou H, et al. Perinatal and neonatal outcomes of 494 babies delivered from 972 vitrified embryo transfers. Fertil Steril. 2012;97(6):1338–42. https://doi.org/10.1016/j.fertnstert.2012.02.051.
Desai NN, Goldstein J, Rowland DY, Goldfarb JM. Morphological evaluation of human embryos and derivation of an embryo quality scoring system specific for day 3 embryos: a preliminary study. Hum Reprod. 2000;15(10):2190–6.
Shimoda Y, Kumagai J, Anzai M, Kabashima K, Togashi K, Miura Y, et al. Time-lapse monitoring reveals that vitrification increases the frequency of contraction during the pre-hatching stage in mouse embryos. J Reprod Dev. 2016;62(2):187–93. https://doi.org/10.1262/jrd.2015-150.
Wells D, Bermudez MG, Steuerwald N, Thornhill AR, Walker DL, Malter H, et al. Expression of genes regulating chromosome segregation, the cell cycle and apoptosis during human preimplantation development. Hum Reprod. 2005;20(5):1339–48. https://doi.org/10.1093/humrep/deh778.
Gardner DK, Lane M, Schoolcraft WB. Physiology and culture of the human blastocyst. J Reprod Immunol. 2002;55(1–2):85–100.
Gardner DK, Balaban B. Assessment of human embryo development using morphological criteria in an era of time-lapse, algorithms and ‘OMICS’: is looking good still important? Mol Hum Reprod. 2016;22(10):704–18. https://doi.org/10.1093/molehr/gaw057.
Graham J, Han T, Porter R, Levy M, Stillman R, Tucker MJ. Day 3 morphology is a poor predictor of blastocyst quality in extended culture. Fertil Steril. 2000;74(3):495–7.
Gardner DK. The impact of physiological oxygen during culture, and vitrification for cryopreservation, on the outcome of extended culture in human IVF. Reprod BioMed Online. 2016;32(2):137–41. https://doi.org/10.1016/j.rbmo.2015.11.008.
Botros L, Sakkas D, Seli E. Metabolomics and its application for non-invasive embryo assessment in IVF. Mol Hum Reprod. 2008;14(12):679–90. https://doi.org/10.1093/molehr/gan066.
Alikani M, Calderon G, Tomkin G, Garrisi J, Kokot M, Cohen J. Cleavage anomalies in early human embryos and survival after prolonged culture in-vitro. Hum Reprod. 2000;15(12):2634–43.
Pereira N, Brauer AA, Melnick AP, Lekovich JP, Spandorfer SD. Prognostic value of growth of 4-cell embryos on the day of transfer in fresh IVF-ET cycles. J Assist Reprod Genet. 2015;32(6):939–43. https://doi.org/10.1007/s10815-015-0478-2.
Zhao P, Li M, Lian Y, Zheng X, Liu P, Qiao J. The clinical outcomes of day 3 4-cell embryos after extended in vitro culture. J Assist Reprod Genet. 2015;32(1):55–60. https://doi.org/10.1007/s10815-014-0361-6.
Cummins JM, Breen TM, Harrison KL, Shaw JM, Wilson LM, Hennessey JF. A formula for scoring human embryo growth rates in in vitro fertilization: its value in predicting pregnancy and in comparison with visual estimates of embryo quality. J In Vitro Fert Embryo Transf. 1986;3(5):284–95.
Kroener LL, Ambartsumyan G, Pisarska MD, Briton-Jones C, Surrey M, Hill D. Increased blastomere number in cleavage-stage embryos is associated with higher aneuploidy. Fertil Steril. 2015;103(3):694–8. https://doi.org/10.1016/j.fertnstert.2014.12.090.
Fenwick J, Platteau P, Murdoch AP, Herbert M. Time from insemination to first cleavage predicts developmental competence of human preimplantation embryos in vitro. Hum Reprod. 2002;17(2):407–12.
Papanikolaou EG, Kolibianakis EM, Tournaye H, Venetis CA, Fatemi H, Tarlatzis B, et al. Live birth rates after transfer of equal number of blastocysts or cleavage-stage embryos in IVF. A systematic review and meta-analysis. Hum Reprod. 2008;23(1):91–9. https://doi.org/10.1093/humrep/dem339.
Glujovsky D, Farquhar C, Quinteiro Retamar AM, Alvarez Sedo CR, Blake D. Cleavage stage versus blastocyst stage embryo transfer in assisted reproductive technology. Cochrane Database Syst Rev. 2016;6:CD002118. https://doi.org/10.1002/14651858.CD002118.pub5.
Tang R, Catt J, Howlett D. Towards defining parameters for a successful single embryo transfer in frozen cycles. Hum Reprod. 2006;21(5):1179–83. https://doi.org/10.1093/humrep/dei490.
Blake DA, Farquhar CM, Johnson N, Proctor M. Cleavage stage versus blastocyst stage embryo transfer in assisted conception. Cochrane Database Syst Rev. 2007;4:CD002118. https://doi.org/10.1002/14651858.CD002118.pub3.
Maheshwari A, Hamilton M, Bhattacharya S. Should we be promoting embryo transfer at blastocyst stage? Reprod BioMed Online. 2016;32(2):142–6. https://doi.org/10.1016/j.rbmo.2015.09.016.
Glujovsky D, Blake D, Farquhar C, Bardach A. Cleavage stage versus blastocyst stage embryo transfer in assisted reproductive technology. Cochrane Database Syst Rev. 2012;7:CD002118. https://doi.org/10.1002/14651858.CD002118.pub4.
Rizos D, Lonergan P, Boland MP, Arroyo-Garcia R, Pintado B, de la Fuente J, et al. Analysis of differential messenger RNA expression between bovine blastocysts produced in different culture systems: implications for blastocyst quality. Biol Reprod. 2002;66(3):589–95.
Kallen B, Finnstrom O, Lindam A, Nilsson E, Nygren KG, Olausson PO. Blastocyst versus cleavage stage transfer in in vitro fertilization: differences in neonatal outcome? Fertil Steril. 2010;94(5):1680–3. https://doi.org/10.1016/j.fertnstert.2009.12.027.
Dar S, Librach CL, Gunby J, Bissonnette F, Cowan L. Increased risk of preterm birth in singleton pregnancies after blastocyst versus day 3 embryo transfer: Canadian ART Register (CARTR) analysis. Hum Reprod. 2013;28(4):924–8. https://doi.org/10.1093/humrep/des448.
Martins WP, Nastri CO, Rienzi L, van der Poel SZ, Gracia C, Racowsky C. Blastocyst vs cleavage-stage embryo transfer: systematic review and meta-analysis of reproductive outcomes. Ultrasound Obstet Gynecol. 2017;49(5):583–91. https://doi.org/10.1002/uog.17327.
Acknowledgements
We greatly appreciate and thank Professor Yuanqing Yao, from the Department of Obstetrics and Gynecology, Chinese PLA General Hospital, for his comments and revision of the manuscript.
Funding
This study was funded by the medical scientific research project of Lanzhou Military Region (project number: CLZ14JB08).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Ethical approval
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent
Informed consent was obtained from all individual participants included in the study.
Conflict of interest
The authors declare that they have no conflict of interest.
Electronic supplementary material
ESM 1
(XLSX 15 kb).
Rights and permissions
About this article
Cite this article
Zhao, X., Ma, B., Mo, S. et al. Improvement of pregnancy outcome by extending embryo culture in IVF-ET during clinical application. J Assist Reprod Genet 35, 321–329 (2018). https://doi.org/10.1007/s10815-017-1065-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10815-017-1065-5