Optimization of ART programs in women of late reproductive age by co-cultivation of oocytes with donor extracellular vesicles from follicular fluid
Akhmedova Z.F., Sysoeva A.P., Gavrilov M.Yu., Zingerenko B.V., Shevtsova Yu.A., Silachev D.N., Ekimov A.N., Makarova N.P., Kulakova E.V., Kalinina E.A.
Background: The effectiveness of infertility treatment using assisted reproductive technologies (ART) decreases with the age of the woman due to the aging of the germ cells. It has been previously shown that co-cultivation of embryos with autologous extracellular vesicles from follicular fluid (EVFF) improves the embryological stage of treatment. However, in women of late reproductive age, biological fluids senesce and lose their effectiveness. Therefore, a hypothesis about the use of donor EVFF to improve the results of ART has been proposed.
Objective: To evaluate the effectiveness of donor EVFF in co-cultivation of embryos of women over 40 years.
Materials and methods: The study was conducted among 79 married infertile couples. Inclusion criteria: the woman's age > 40 years, normal karyotype both in a woman and a man. The patients were divided into two groups: the main group (n=40) with co-cultivation of oocytes with donor EVFF and the comparison group (n=39) with standard cultivation. Stimulation of ovarian function was performed according to the protocol with gonadotropin-releasing hormone antagonists. Transvaginal puncture was done under anesthesia. In group 1, donor EVFF were added to fertilized oocytes. The next day, they were transferred to fresh culture medium. The main evaluation parameters included: fertilization rate, blastocyst formation, and clinical pregnancy.
Results: In the group with donor EVFF, the fertilization rate was higher (84.8% vs. 70%, p=0.008), the proportion of embryo transfer cancellations was lower (22.5% vs. 46.1%, p=0.021). The frequency of pregnancy and genetically normal embryos did not statistically differ.
Conclusion: Donor EVFF improve embryological parameters but do not affect clinical outcomes in women of late reproductive age.
Authors’ contributions: Akhmedova Z.F. – study concept, data collection, data processing, text composition; Sysoeva A.P. – conduction of embryo stage of infertility treatment, critical re-evaluation of obtained results; Gavrilov M.Yu. – conduction of embryo stage of infertility treatment, extracellular vesicles identification; Zingerenko B.V. – conduction of embryo stage of infertility treatment, data processing; Shevtsova Yu.A. – extracellular vesicles identification using ultracentrifugation; Silachev D.N. – critical re-evaluation of the manuscript and the obtained results; Ekimov A.N. – preimplantation genetic testing conduction; Makarova N.P. – study concept, data processing, text composition; Kulakova E.V. – critical re-evaluation of obtained results from the point of obstetrics and gynecology; Kalinina E.A. – manuscript final approval.
Conflicts of interest: The authors declare no conflicts of interest in connection with the publication of this article.
Funding: The study was carried out within the initiative scientific project “Study of the influence of extracellular vesicles from reproductive organs and tissues biological fluids on gametes, the process of fertilization and early human embryogenesis and implantation” (2025–2027, supervisor — N.P. Makarova), Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia.
Ethical Approval: The study was approved by the bioethics committee of the Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia (Protocol #10 of October 20, 2022).
Patient Consent for Publication: All patients provided informed consent for the publication of their data.
Authors' Data Sharing Statement: The data supporting the findings of this study are available on request from the corresponding author after approval from the principal investigator.
For citation: Akhmedova Z.F., Sysoeva A.P., Gavrilov M.Yu., Zingerenko B.V., Shevtsova Yu.A., Silachev D.N., Ekimov A.N., Makarova N.P., Kulakova E.V., Kalinina E.A.. Optimization of ART programs in women of late reproductive age by co-cultivation of oocytes with donor extracellular vesicles from follicular fluid.
Akusherstvo i Ginekologiya/Obstetrics and Gynecology. 2025; (8): 133-142 (in Russian)
https://dx.doi.org/10.18565/aig.2025.104
Keywords
ART
infertility
late reproductive age
co-cultivation
follicular fluid
extracellular vesicles
IVF
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Received 17.04.2025
Accepted 11.08.2025
About the Authors
Zumriiat F. Akhmedova, PhD student at the Prof. B.V. Leonov Department for Assisted Technologies in Infertility Treatment, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, zyuka-1997@mail.ru,https://orcid.org/0000-0002-4483-8820
Anastasia P. Sysoeva, PhD, Clinical Embryologist at the Prof. B.V. Leonov Department for Assisted Technologies in Infertility Treatment, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, a_sysoeva@oparina4.ru, https://orcid.org/0000-0002-6502-4498
Maxim Yu. Gavrilov, Junior Researcher at the Prof. B.V. Leonov Department of Assistive Technologies in Infertility Treatment, Academician V.I. Kulakov National
Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, maxgavr67@gmail.com, https://orcid.org/0000-0001-6189-0287
Boris V. Zingerenko, Junior Researcher at the Prof. B.V. Leonov Department of Assistive Technologies in Infertility Treatment, Academician V.I. Kulakov National
Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, b_zingerenko@oparina4.ru, https://orcid.org/0000-0002-8784-5502
Yulia A. Shevtsova, Junior Researcher at the Cell Technologies Laboratory, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, u_shevtsova@oparina4.ru
Denis N. Silachev, Dr. Bio. Sci., Head of the Cell Technologies Laboratory, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, d_silachev@oparina4.ru, https://orcid.org/0000-0003-0581-9755
Alexey N. Ekimov, PhD, Head of the Laboratory of Preimplantation Genetic Testing and Genetic Diagnostics, Academician V.I. Kulakov National Medical Research Center
for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, a_ekimov@oparina4.ru,
https://orcid.org/0000-0001-5029-0462
Natalya P. Makarova, Dr. Bio. Sci., Leading Researcher at the Prof. B.V. Leonov Department of Assistive Technologies in Infertility Treatment, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4,
np_makarova@oparina4.ru, https://orcid.org/0000-0003-1396-7272
Elena V. Kulakova, Dr. Med. Sci., Senior Researcher at the Prof. B.V. Leonov Department of Assistive Technologies in Infertility Treatment, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, e_kulakova@oparina4.ru, https://orcid.org/0000-0002-4433-4163
Elena A. Kalinina, Dr. Med. Sci., Professor, Head of the Prof. B.V. Leonov Department of Assistive Technologies in Infertility Treatment, Academician V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia, 117997, Russia, Moscow, Ac. Oparin str., 4, e_kalinina@oparina4.ru, https://orcid.org/0000-0002-8922-2878
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