Predictors of spontaneous human blastocyst hatching in ART programs


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The spontaneous hatching inability of human blastocysts in ART programs may lead to failure of implantation and to the lack of pregnancy. The mechanism of spontaneous hatching and the predictors of its efficiency are not fully known and require further study. Objective: To investigate the role of cellular, genetic, clinical, laboratory, and iatrogenic factors in the efficiency of spontaneous hatching of human blastocysts in ART programs. Material and methods. The investigation enrolled 83 human blastocysts donated for researches. Light microscopy was used to determine the degree of maturity and quality of oocytes and embryos and the thickness of the pellucid zone, to reveal oocyte dysmorphisms, and to monitor hatching. The expression of CTSL-2, GATA3, and CGB mRNA was estimated using a real-time polymerase chain reaction assay. Results. The state of the pellucid zone and the quality of the gametes that were employed to derive blastocysts had no significant effect on hatching with the exception of extracytoplasmic dysmorphisms of the oocytes, in the presence of which the hatching efficiency increased 4.3-fold. The values of all three components of the Gardner’s blastocyst grade were significantly higher in the effective hatching group. The hatching success was influenced by the lower level of follicle-stimulating hormone (FSH), by superovulation stimulated with recombinant FSH, and by the use of GnRH agonists as an ovulation trigger. The effectively hatched blastocyst group displayed a higher expression of CTSL2, GATA3, and CGB mRNA. Embryo maturity and quality were associated with the CTSL2 and GATA3 mRNA expression that was higher in developmental stages 5-6 embryos and in class A embryos. Conclusion. The efficiency of spontaneous human blastocyst hatching is mainly affected by the quality of blastocysts themselves rather than by that of the pellucid zone and gametes. The blastocyst can model its further development through its own genetic factors. The expression of CTSL2, GATA3, and CGB mRNA is higher in best-quality blastocysts, which allows them to spontaneously hatch. The choice of a superovulation protocol plays a role in the efficiency of hatching.

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Nataliya Dolgushina

Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: n_dolgushina@oparina4.ru
M.D., Ph.D., M.P.H., Head of R&D Department

Espet Ibragimova

Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: Espet2007@yandex.ru
PhD student of ART Department

Andrey Romanov

Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: romanov1553@yandex.ru
clinical resident

Olga Burmenskaya

Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: o_bourmenskaya@oparina4.ru
PhD, DSc, Senior Researcher of Molecular-Genetics Department

Nataliya Makarova

Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: np_makarova@oparina4.ru
PhD, Researcher of ART Department

Ramin Shafei

Lomonosov Moscow State University

Email: shafei@mail.ru
PhD, Leading Researcher, Biological Faculty, Dept. Embryology.

Anastasiya Syrkasheva

Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: anast.syrkasheva@gmail.com
M.D., Ph.D., researcher of ART Department

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