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Research Article Human Reproduction

Laser collapse performance in blastocysts applied before vitrification

José Fernando de Macedo, Maristela Rodrigues Oliveira, Luiz Mauro Oliveira Gomes, Gustavo Capinzaiki de Macedo, Giovanna Capinzaiki de Macedo, Daniela Oliveira Gomes, Sandra Irene Sprogis dos Santos

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The objetive of the present study was to investigate the effect of the reduction of blastocele fluid before vitrification with the application of collapsing and the effects of this procedure on the expected results of in vitro fertilization protocols. Methods: A retrospective casecontrol study was conducted with patients who sought assisted fertilization services from January 2015 to October 2017 at Reproferty Clinic. After submission to the ethics committee, data from 106 patients were divided into two groups: 42 cases represented by those who had artificially collapsed blastocysts per laser pulse before vitrification and 64 controls corresponding to patients where artificial shrinkage of expanded blastocysts was not performed. By statistical analysis, the means and standard deviation of each group were obtained, which were compared by the Mann-Whitney test, at a 95% significance level. Results: the results showed that there was statistical significance comparing the group of cases and controls in the parameters related to embryo freezing, survival and transference as well as to the rates of positive beta human chorionic gonadotropin, clinical pregnancy and live births. Conclusions: the study demonstrated evidence that vitrification after artificial laser collapse of the blastocyst is safe, effective, and more efficient than vitrification of non-collapsed blastocysts in vitro fertilization protocols, however further studies are needed to evaluate the efficiency of artificial shrinkage in routine protocols.


blastocyst; laser collapse; vitrification; in vitro fertilization; embryo implantation; artificial shrinkage


1. Roque M, Valle M, Kostolias A, Sampaio M, Geber S. Freeze-all cycle in reproductive medicine: current perspectives. JBRA Assist Reprod. 2017;21(1):49-53. PMid:28333033.

2. Evans J, Hannan NJ, Edgell TA, Vollenhoven BJ, Lutjen PJ, Osianlis T, et al. Fresh versus frozen embryo transfer: backing clinical decisions with scientific and clinical evidence. Hum Reprod Update. 2014;20(6):808-21. PMid:24916455.

3. Liebermann J, Pelts EJ, Matthews JM, Sanchez SR, Rapisarda J, Lederer K. Does artificial collpase of human day 6 blastocysts prior to the cooling steps of vitrification improve their probability of increased outcome? Fertil Steril. 2012;98(Suppl 3):S127.

4. Al-Azawi T, Tavukcuoglu S, Khaki AA, Al-Hasani S. Cryopreservation of human oocytes, zygotes, embryos and blastocysts: a comparison study between slow freezing and ultra rapid (vitrification) methods. Middle East Fertil Soc J. 2013;18(4):223-32.

5. Ortega-Hrepich C, Stoop D, Guzmán L, Van Landuyt L, Tournaye H, Smitz J, et al. A “freeze-all” embryo strategy after in vitro maturation: a novel approach in women with polycystic ovary syndrome? Fertil Steril. 2013;100(4):1002-7. PMid:23850301.

6. Cota AMM, Lages EL, Mendonça HC, Silva ALM, Amaral MCMS, Caetano JPJ. Avaliação dos resultados de vitrificação de blastocistos. JBRA Assist Reprod. 2011;15(4):27-30.

7. Blockeel C, Drakopoulos P, Santos-Ribeiro S, Polyzos NP, Tournaye H. A fresh look at the freeze-all protocol: a SWOT analysis. Hum Reprod. 2016;31(3):491-7. PMid:26724793.

8. Dieamant FC, Petersen CG, Mauri AL, Comar V, Mattila M, Vagnini LD, et al. Fresh embryos versus freeze-all embryos - transfer strategies: nuances of a meta-analysis. JBRA Assist Reprod. 2017;21(3):260-72. PMid:28837037.

9. Roque M, Valle M, Guimarães F, Sampaio M, Geber S. Freeze-all policy: fresh vs. frozen-thawed embryo transfer. Fertil Steril. 2015;103(5):1190-3. PMid:25747130.

10. Elnahas A, Alcolak E, Marar EA, Elnahas T, Elnahas K, Palapelas V, et al. Vitrification of human oocytes and different development stages of embryos: an overview. Middle East Fertil Soc J. 2010;15(1):2-9.

11. Rezazadeh Valojerdi 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(6):347-54. PMid:19513822.

12. Darwish E, Magdi Y. Artificial shrinkage of blastocoel using a laser pulse prior to vitrification improves clinical outcome. J Assist Reprod Genet. 2016;33(4):467-71. PMid:26843389.

13. Liebermann J. (2012). Vitrification of oocytes and embryos. In: Katkov I, editor. Current frontiers in cryobiology. Rijeka: InTech. p. 169-184.

14. Shetye-Kapdi S, Kagalwale S, Ghandi GN, Pangerkar P. Slow-rate freezing versus vitrification for oocytes and embryos: an evaluation In: Allahbadia GN, Chawla MM, Das RB, Garcia EV, Gandhi G, Merchant R, editors. The Art & Science of Assisted Reproductive Techniques (ART). London: Taylor & Francis; 2004. p. 605.

15. Liebermann J, Tucker MJ. Effect of carrier system on the yield of human oocytes and embryos as assessed by survival and developmental potential after vitrification. Reproduction. 2002;124(4):483-9. PMid:12361466.

16. Liebermann J, Tucker MJ. Comparison of vitrification and conventional cryopreservation of day 5 and day 6 blastocysts during clinical application. Fertil Steril. 2006;86(1):20-6. PMid:16762345.

17. Stachecki JJ, Willadsen SM, Wiemer K, Garrisi J, Cohen J. S3 vitrification: a safe, simple, and successful method for blastocyst vitrification. Fertil Steril. 2007;88:S347-8.

18. Desai N, Szeptycki J, Scott M, AbdelHafez FF, Goldfarb J. Artificial collapse of blastocysts before vitrification: mechanical vs. laser technique and effect on survival, cell number, and cell death in early and expanded blastocysts. Cell Preserv Technol. 2008;6(3):181-90.

19. Mukaida T, Oka C, Goto T, Takahashi K. Artificial shrinkage of blastocoeles using either a micro-needle or a laser pulse prior to the cooling steps of vitrification improves survival rate and pregnancy outcome of vitrified human blastocysts. Hum Reprod. 2006;21(12):3246-52. PMid:16936299.

20. Fischer E, Givens C, Conaghan J. Optimization of blastocyst vitrification may require artificial colapse. Fertil Steril. 2011;95(4):S5-6.

21. Bodri D, Sugimoto T, Yao Serna J, Kawachiya S, Kato R, Matsumoto T. Blastocyst collapse is not an independent predictor of reduced live birth: a time-lapse study. Fertil Steril. 2016;105(6):1476-83. PMid:26940789.

22. Taylor TH, Gilchrist JW, Hallowell SV, Hanshew KK, Orris JJ, Glassner MJ, et al. The effects of different laser pulse lengths on the embryo biopsy procedure and embryo development to the blastocyst stage. J Assist Reprod Genet. 2010;27(11):663-7. PMid:20686919.

23. Iriarte YF, Morrilo MJL, Lizana EO, Laso A. Artificial collapse of human blastocysts, before vitrification, improves survival and pregnancy rates. Reprod Biomed Online. 2012;24(Suppl 1):S9.

24. Kazemi P, Dashtizad M, Shamsara M, Mahdavinezhad F, Hashemi E, Fayazi S, et al. Effect of blastocoel fluid reduction before vitrification on gene expression in mouse blastocysts. Mol Reprod Dev. 2016;83(8):735-42. PMid:27409768.

25. Kader AA, Choi A, Orief Y, Agarwal A. Factors affecting the outcome of human blastocyst vitrification. Reprod Biol Endocrinol. 2009;7(1):99. PMid:19758458.

26. Vanderzwalmen P, Connan D, Grobet L, Wirleitner B, Remy B, Vanderzwalmen S, et al. Lower intracellular concentration of cryoprotectants after vitrification than after slow freezing despite exposure to higher concentration of cryoprotectant solutions. Hum Reprod. 2013;28(8):2101-10. PMid:23592220.

27. Mukaida T, Nakamura S, Tomiyama T, Wada S, Oka C, Kasai M, et al. Vitrification of human blastocysts using cryoloops: clinical outcome of 223 cycles. Hum Reprod. 2003;18(2):384-91. PMid:12571178.

28. Duke M, Basile N, Johnson M, Copperman A, Sandler B, Barritt J. Hatched blastocysts that collapse during slow freezing have a higher post-thaw survival rate. Fertil Steril. 2004;82(Suppl 2):S150-1.

29. Son W-Y, Yoon S-H, Yoon H-J, Lee S-M, Lim J-H. Pregnancy outcome following transfer of human blastocysts vitri®ed on electron microscopy grids after induced collapse of the blastocoele. Hum Reprod. 2003;18(1):137-9. PMid:12525454.

30. Kovačič B, Taborin M, Vlaisavljević V. Artificial blastocoel collapse of human blastocysts before vitrification and its effect on re-expansion after warming - a prospective observational study using time-lapse microscopy. Reprod Biomed Online. 2018;36(2):121-9. PMid:29212605.

31. Fields RA, Werland HJ, Nguyen J, Sieren KR, Turner TG, Silverberg KM. Laser collapsing of blastocysts prior to vitrification leads to better embryonic survival and improved overall IVF cycle outcome. Fertil Steril. 2013;100(3):S106.

32. Marcos J, Pérez-Albalá S, Mifsud A, Molla M, Landeras J, Meseguer M. Collapse of blastocysts is strongly related to lower implantation success: a time-lapse study. Hum Reprod. 2015;30(11):2501-8. PMid:26355116.

33. Macer ML, Barritt J, Surrey MW, Danzer H, Ghadir S, Chang W, et al. Pregnancy outcomes following single versus double euploid embryo transfer. Fertil Steril. 2017;107(3):e28-9.

34. Chae SJ, Hur CY, Lee WD, Lim JH. Single embryo transfer versus double embryo transfer in vitrified frozen-thawed blastocyst transfer cycles: retrospective analysis of 967 cryopreserved cycles with no more than two blastocysts. Fertil Steril. 2008;90(Suppl 1):S366.

35. Gremeau AS, Brugnon F, Bouraoui Z, Peikhrishvili R, Janny L, Pouly JL. Outcome of elective single or double embryo transfer in first and second IVF/ICSI cycles. Gynecol Obstet Fertil. 2011;39(2):70-5. PMid:21316284.

36. Kosmas I, Van der Elst J, Devroey P, Tournaye H. Elective single embryo transfer. BJOG. 2008;10:163-70.

37. Forman EJ, Hong KH, Ferry KM, Tao X, Taylor D, Levy B, et al. In vitro fertilization with single euploid blastocyst transfer: a randomized controlled trial. Fertil Steril. 2013;100(1):100-7. PMid:23548942.

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