Co-cultivation of mastocytes and cumulus cells in polycystic ovary syndrome

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Abstract

The purpose of the study was to study the cytokine-producing function of cumulus cells of the ovaries in PCOS during co-cultivation of cumulus cells with mast cells to identify their mutual influence in the immunopathogenesis of PCOS.

The study was approved by the Interdisciplinary Ethics Committee of the FSBEI of HE of the Ministry of Health of the Russian Federation. A permanent human mast cell line HMC-1 (Human mast cell, ATCC, USA) and a primary culture of cumulus cells were used. The condition of cumulus cells and mastocytes was determined using fluorescent dyes followed by flow cytometry before co-cultivation and after 7 days co-cultivation. The levels of IL-6, IL-10, and IFNã were studied on days 1, 3, and 7 of the experiment.

In monoculture and during co-culture of cells, the synthesis of cytokines IL-6, IL-10, and IFNã continues, but to varying degrees of severity. In the permanent mast cell line, an increase in IL-6, IL-10, and IFNã is observed on the 1st, 3rd, and 7th days of cultivation. On the first day, cytokine levels of donor cumulus cells and mastocytes did not differ significantly (p > 0.05). In the culture of donor cumulus cells, the synthesis of IL-6, IL-10, and IFNã progressively increases by the 7th day of the experiment (p < 0.05). On the first day of the experiment, the balance of Th1/Th2 cytokines in PCOS was twice as high as in healthy women. The ratio increased, and on the 7th day reached 3.83 times higher than the ratio of Th1/Th2 cytokines in the control group. Hyperproduction of IFNã by mastocytes was most significant when they were co-cultured with cumulus cells, in monoculture mast cells synthesized excessively cytokine twice from the initial values of IFNã, and the monoculture cumulus cells in PCOS it practically did not contain.

The studied cytokines are regulators of the function of ovarian cumulus cells and factors that increase the competence of the oocyte, indicating the need for their correction, which will allow a real influence on the links of pathogenesis in PCOS in the future.later on.

About the authors

O. V. Valikova

Pacific State Medical University; Regional Clinical Hospital No. 2; Cliniс “Plastic Surgery”

Author for correspondence.
Email: renalex.99@mail.ru

Junior Research Associate, Endocrinologist

Russian Federation, Vladivostok; Vladivostok; Vladivostok

V. V. Zdor

Pacific State Medical University; Cliniс for Diabetes and Endocrine Diseases

Email: renalex.99@mail.ru

PhD, MD (Medicine), Leading Research Associate, Endocrinologist

Russian Federation, Vladivostok; Vladivostok

Ya. N. Tikhonov

Pacific State Medical University

Email: renalex.99@mail.ru

Senior Lecturer, Department of Pathological Anatomy

Russian Federation, Vladivostok

A. V. Boroda

A. Zhirmunsky National Scientific Center of Marine Biology, Far East Branch, Russian Academy of Sciences

Email: renalex.99@mail.ru

PhD, Senior Research Associate, Laboratory of Cell Technology, Deputy Director for Scientific Activities

Russian Federation, Vladivostok

K. G. Kolbin

Clinic for Reproduction and Genetics “Next Generation Clinic”

Email: renalex.99@mail.ru

PhD (Biology), Embryologist

Russian Federation, Vladivostok

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Supplementary files

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2. Figure 1. Cytokine levels during co-cultivation of a permanent line of human mast cells HMC-1 and cumulus cells of patients with polycystic ovary syndrome

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3. Figure 2. Cytokine levels during co-cultivation of a permanent line of human mast cells HMC-1 and cumulus cells of healthy donors

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4. Figure 3. Cytometry parameters in the co-cultivation of a permanent line of human mast cells HMC-1 and cumulus cells of healthy donors

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5. Figure 4. Cytometry parameters in the joint cultivation of a permanent line of human mast cells HMC-1 and cumulus cells of patients with polycystic ovary syndrome

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Copyright (c) 2024 Valikova O.V., Zdor V.V., Tikhonov Y.N., Boroda A.V., Kolbin K.G.

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