Granulosa Cells Morphological Changes in Infertile Women with PCOS Compared to Healthy Subjects Under-going IVF

Publish Year: 1398
نوع سند: مقاله کنفرانسی
زبان: English
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RROYAN20_317

تاریخ نمایه سازی: 29 مهر 1398

Abstract:

Background: During follicular development, intraovarian par-acrine signals coordinate granulosa cells (GCs) proliferation, theca cell differentiation, and oocyte maturation. In women with polycystic ovary syndrome (PCOS), ovarian hyperandrogenism and altered intrafollicular paracrine signaling perturb follicular survival, growth, and selection, causing accumulation of small antral follicles within the periphery of the ovary. Morphological Changes of (GCs) have not been thoroughly investigated during this syndrome.Materials and Methods: The study was performed on ovarian GCs of 220 women who referred for IVF-ET. The PCOS group consisted of 120 PCOS patients, and 100 women with healthy ovaries were considered as the normal group. The syndrome was diagnosed by a gynecologist according to Rotterdam cri-teria (Rotterdam ESHRE/ASRM, 2004). After washing and trypsinization with 0.25% trypsin/EDTA solution, isolated GCs were resuspended in DMEM/F12 culture medium, and counted by a hemocytometer. Trypan blue 0.4% was used to determine the viable and total cell counts. GCs were seeded in 6-well plates (2×106 cells/well) and cultivated in 3 mL DMEM/F12 supplemented with 5% Fetal Bovine Serum, 1% L-Glutamine, and 1% Penicillin/ Streptomycin for 48 hours. In order to evalu-ate the purity, morphology, and growth rate of GCs, cell culture plates photography was done, and images were analyzed by Im-ageJ software.Results: Typical morphology of normal GCs (N-GCs) was spher-ical. However, spherical and non-spherical cells were observed in both groups, but the ratio of spherical to non-spherical PCO-GCs was significantly (P<0.0001) lower than that N-GCs (0.36 ± 0.02 versus 7.34±0.38). Photographs analysis showed that the mean value of the diameter of spherical PCO-GCs were significantly (P<0.0001) smaller than N-GCs (6.022 ± 0.054 versus 10.21 ± 0.058 μm). Trypan blue exclusion test showed significant differ-ence in viable GCs percent between PCOS patients and normal women (72.93 ± 2.12 % versus 83.36 ± 2.42 %).Conclusion: The current study findings suggest that metabol-ic changes in GCs of women with PCOS undergoing IVF-ET correlate with significant morphological abnormalities and de-creased viability. Due to bidirectional communication between GCs with oocyte, the quality of the oocyte, consequently, the quality of the embryo are adversely affected.

Authors

F Masjedi

Department of Physiology, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran

S Keshtgar

Department of Physiology, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran