Predictors of spontaneous human blastocyst hatching in ART programs
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.Dolgushina N.V., Ibragimova E.O., Romanov A.Yu., Burmenskaya O.V., Makarova N.P., Shafei R.A., Syrkasheva A.G.
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.
Keywords
Supplementary Materials
- Figure. Evaluation of embryos according to Gardner's classification in the effective (Hatching +) and inefficient (Hatching-) spontaneous hatching groups: A - in groups according to the degree of maturity of the blastocyst (5, 4, 2-3); B - on the quality of intracellular mass (A, B and C); C - on the quality of trophectoderm (A, B and C)
- Table 1. Characteristics of blastocysts who have committed and not committed spontaneous hatching
- Table 2. Dependence of the expression of embryonic genetic factors and the ability of the blastocyst to spontaneous hatching
- Table 3. Dependence of expression of embryonic genetic factors on blastocyst quality
References
1. Bissonnette F., Cohen J., Collins J., Cowan L., Dale S., Dill S. et al. Incidence and complications of multiple gestation in Canada: proceedings of an expert meeting. Reprod. Biomed. Online. 2007; 14(6): 773-790.
2. Gunby J., Daya S. Assisted reproductive technologies (ART) in Canada: 2002 results from the Canadian ART Register. Fertil. Steril. 2006; 86(5): 1356-64.
3. Кузьмичев Л., Смольникова В., Калинина Е., Дюжева Е. Принципы комплексной оценки и подготовки эндометрия у пациенток программ вспомогательных репродуктивных технологий. Акушерство и гинекология. 2010; 5: 32-6. [Kuzmichev L.N., Smolnikova V.Yu., Kalinina Ye.A., Dyuzheva Ye. V. The principles of complex evaluation and preparation of the endometrium in patients of assisted reproductive technology programs. Akusherstvo i Ginekologiya/Obstetrics and Gynecology. 2010; 5: 32-6. (in Russian)]
4. Ковальская Е.В., Сыркашева А.Г., Романов А.Ю., Макарова Н.П., Долгушина Н.В. Современные представления о компактизации эмбрионов человека в условиях in vitro. Технологии живых систем. 2017; 1: 25-35. [Kovalskaya E.V., Syrkasheva A.G., Romanov A.Yu., Makarova N.P., Dolgushina N.V. Modern ideas about the compaction of human embryos in vitro. Tehnologii zhivyih sistem. 2017; 1: 25-35. (in Russian)]
5. Wassarman P. Zona pellucida glycoproteins. J. Biol. Chem. 2008; 283(36): 24285-9.
6. Шафеи Р.А., Сыркашева А.Г., Романов А.Ю., Макарова Н.П., Долгушина Н.В., Семенова М.Л. Хетчинг бластоцисты у человека. Онтогенез. 2017; 48(1): 8-20. [Shafei R.A., Syrkasheva A.G., Romanov A.Yu., Makarova N.P., Dolgushina N.V., Semenova M.L. Hatching blastocysts in humans. Ontogenez. 2017; 48 (1): 8-20. (in Russian)]
7. Sathananthan H., Menezes J., Gunasheela S. Mechanics of human blastocyst hatching in vitro. Reprod. Biomed. Online. 2003; 7(2): 228-34.
8. Quesada V., Sanchez L.M., Alvarez J., Lopez-Otin C. Identification and characterization of human and mouse ovastacin: a novel metalloproteinase similar to hatching enzymes from arthropods, birds, amphibians, and fish. J. Biol. Chem. 2004; 279(25): 26627-34.
9. Гоголевский П.А. Вспомогательный хэтчинг: показания, применение, результаты (обзор литературы). Проблемы репродукции. 1998; 4(1): 10-3. [Gogolevsky P.A. Auxiliary hatching: indications, application, results (literature review). Problemy reproduktsii. 1998; 4(1): 10-3. (in Russian)]
10. Seshagiri P.B., Sen Roy S., Sireesha G., Rao R.P. Cellular and molecular regulation of mammalian blastocyst hatching. J. Reprod. Immunol. 2009;83(1-2): 79-84.
11. Ибрагимова Э.О., Долгушина Н.В., Сыркашева А.Г., Романов А.Ю., Языкова О.И., Макарова Н.П. Роль вспомогательного хетчинга в программах лечения бесплодия методами вспомогательных репродуктивных технологий: обзор литературы. Гинекология. 2016; 18(2): 44-7. [Ibragimova E.O., Dolgushina N.V., Syrkasheva A.G., Romanov A.Yu., Yazykova O.I., Makarova N.P. The role of auxiliary hatching in infertility treatment programs by assisted reproductive technologies: a literature review. Ginekologiya. 2016; 18 (2): 44-7. (in Russian)]
12. Menezes J., Gunasheela S., Sathananthan H. Video observations on human blastocyst hatching. Reprod. Biomed. Online. 2003; 7(2): 217-8.
13. Cooper T.G., Noonan E., von Eckardstein S., Auger J., Baker H.W., Behre H.M. et al. WHO reference values for human semen characteristics. Hum. Reprod. Update. 2010; 16(3): 231-45.
14. Hammadeh M.E., Fischer-Hammadeh C., Ali K.R. Assisted hatching in assisted reproduction: a state of the art. J. Assist. Reprod. Genet. 2011; 28(2): 119-28.
15. Rienzi L., Ubaldi F., Iacobelli M., Romano S., Minasi M.G., Ferrero S. et al. Significance of morphological attributes of the early embryo. Reprod. Biomed. Online. 2005; 10(5): 669-81.
16. Sirard M.A., Richard F., Blondin P., Robert C. Contribution of the oocyte to embryo quality. Theriogenology. 2006; 65(1): 126-36.
17. Долгушина Н.В., Сокур С.А., Глинкина Ж.И., Калинина Е.А. Исходы программ вспомогательных репродуктивных технологий у супружеских пар с различными видами патозооспермии у мужчин. Акушерство и гинекология. 2013; 10: 69-75. [Dolgushina V.F., Sokur S.A., Glinkina Zh.I., Kalinina E.A. Outcomes of assisted reproductive technology programs in married couples with different types of pathozoospermia in men. Akusherstvo i ginekologiya/Obstetrics and Gynecology. 2013; 10: 69-75. (in Russian)]
18. Сыркашева А.Г., Казакова В.В., Долгушина Н.В., Романов А.Ю., Андреева М.Г., Яроцкая Е.Л. Реализация программ вспомогательных репродуктивных технологий у пациенток с агрегатами гладкого эндоплазматического ретикулума в цитоплазме ооцитов. Акушерство и гинекология. 2016; 7: 54-9. http://dx.doi.org/10.18565/aig.2016.7.54-59 [Syrkasheva A.G., Kazakova V.V., Dolgushina N.V., Romanov A.Yu., Andreeva M.G., Yarotskaya E.L. Implementation of assisted reproductive technology programs in patients with smooth endoplasmic reticulum aggregates in the cytoplasm of oocytes. Akusherstvo i ginekologiya/Obstetrics and Gynecology. 2016; (7): 54-9. (in Russian) http://dx.doi.org/10.18565/aig.2016.7.54-59]
19. Горшкова А.Г., Долгушина Н.В., Макарова Н.П., Ковальская Е.В., Калинина Е.А. Факторы риска развития дисморфизмов ооцитов в программах вспомогательных репродуктивных технологий. Акушерство и гинекология. 2015; 5: 66-73. [Gorshkova A.G., Dolgushina N.V., Makarova N.P., Kovalskaya E.V., Kalinina E.A. Risk factors for oocyte dysmorphisms in assisted reproductive technology programs. Akusherstvo i ginekologiya/Obstetrics and Gynecology. 2015; 5: 66-73. (in Russian)]
20. Sireesha G., Mason R.W., Hassanein M., Tonack S., Navarrete Santos A., Fischer B., Seshagiri P.B. Role of cathepsins in blastocyst hatching in the golden hamster. Mol. Hum. Reprod. 2008; 14(6): 337-46.
21. Adjaye J. Whole-genome approaches for large-scale gene identification and expression analysis in mammalian preimplantation embryos. Reprod. Fertil. Dev. 2005; 17(1-2): 37-45.
22. Home P., Ray S., Dutta D., Bronshteyn I., Larson M., Paul S. GATA3 is selectively expressed in the trophectoderm of peri-implantation embryo and directly regulates Cdx2 gene expression. J. Biol. Chem. 2009; 284(42): 28729-37.
23. Ray S., Dutta D., Rumi M.A., Kent L.N., Soares M.J., Paul S. Context-dependent function of regulatory elements and a switch in chromatin occupancy between GATA3 and GATA2 regulate Gata2 transcription during trophoblast differentiation. J. Biol. Chem. 2009; 284(8): 4978-88.
24. He B., Junping C., Li H., Weihong T., Lintao X., Shikai W. Effects of human menopausal gonadotropin on zona pellucida and pregnancy outcomes of ovarian stimulation protocols. Iran. J. Reprod. Med. 2015; 13(6):337-44.
25. Balakier H., Sojecki A., Motamedi G., Bashar S., Mandel R., Librach C. Is the zona pellucida thickness of human embryos influenced by women’s age and hormonal levels? Fertil. Steril. 2012; 98(1): 77-83.
Received 17.07.2017
Accepted 22.09.2017
About the Authors
Dolgushina Nataliya Vitalievna, M.D., Ph.D., M.P.H., Head of R&D Department, Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia. 117997, Russia, Moscow, Ac. Oparina str. 4. Tel.: +74954384977. E-mail: n_dolgushina@oparina4.ruIbragimova Espet Omarbekovna, PhD student of ART Department, Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia.
117997, Russia, Moscow, Ac. Oparina str. 4. Tel.: +74954382501. E-mail: Espet2007@yandex.ru
Romanov Andrey Yurievich, clinical resident, Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia.
117997, Russia, Moscow, Ac. Oparina str. 4. Tel.: +79031589400. E-mail: romanov1553@yandex.ru
Burmenskaya Olga V., PhD, DSc, Senior Researcher of Molecular-Genetics Department, Research Center of Obstetrics, Gynecology and Perinatology,
Ministry of Health of Russia. 117997, Russia, Moscow, Ac. Oparina str. 4. Tel.: +74954382292. E-mail: o_bourmenskaya@oparina4.ru
Makarova Nataliya Petrovna, PhD, Researcher of ART Department, Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia.
117997, Russia, Moscow, Ac. Oparina str. 4. E-mail: np_makarova@oparina4.ru
Shafei Ramin Akhmedovich, PhD, Leading Researcher, Lomonosov Moscow State University, Biological Faculty, Dept. Embryology.
119234, Russia, Moscow, Leninskie Gory 1-12. Tel.: +74959393900. E-mail: shafei@mail.ru
Syrkasheva Anastasiya Grigorievna, M.D., Ph.D., researcher of ART Department, Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia. 117997, Russia, Moscow, Ac. Oparina str. 4. Tel.: +74954382501. E-mail: anast.syrkasheva@gmail.com
For citations: Dolgushina N.V., Ibragimova E.O., Romanov A.Yu., Burmenskaya O.V., Makarova N.P., Shafei R.A., Syrkasheva A.G. Predictors of spontaneous human blastocyst hatching in ART programs. Akusherstvo i Ginekologiya/Obstetrics and Gynecology. 2018; (2): 88-95. (in Russian)
https://dx.doi.org/10.18565/aig.2018.2.88-95