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. 2023 Nov 29:12:259.
doi: 10.4103/abr.abr_151_23. eCollection 2023.

The Effect of Normal Follicular Fluid on the Differentiation of PCOS Ovarian Stem Cells into Oocyte-Like Cells

Mahin Taheri Moghadam  1   2 , Hanan Nazayer  2 , Saeed Azandeh  1   2 , Ali R Eftekhari Moghadam  3 , Roshan Nikbakht  4
Affiliations

The Effect of Normal Follicular Fluid on the Differentiation of PCOS Ovarian Stem Cells into Oocyte-Like Cells

Mahin Taheri Moghadam et al. Adv Biomed Res. .

Abstract

Background: Polycystic ovarian syndrome (PCOS) is one of the causes of infertility for which treatment methods do not have a high rate of pregnancy. In this study, the stem cells in the follicular fluid (FF) of patients were grown in the normal FF, and their differentiation into oocytes was evaluated.

Materials and methods: The FF of PCOS patients was centrifuged, and their cells were cultured with and without 20% normal FF for 2 weeks. The cells were evaluated for their morphology by inverted microscope and for markers of stem cells (NANOG and OCT4) and oocytes (zona pellucida (ZP) 2 and ZP3) by RT-PCR and immunocytochemistry. The amount of steroids was measured by enzyme-linked immunosorbent assay (ELISA).

Results: The cells were all round on day 0. After that, they had a heterogeneous morphology (fibroblast-like cells, epithelial-like cells, and round oocyte-like cells). In the first week, NANOG and OCT4 genes in the study group were less expressed than those in the control group (P < 0.0001) (~0.5-fold), while ZP2 and Z3 genes were more expressed (P < 0.0001) (~2-fold). In the second week, stem cell genes were more expressed in the control group (~2 fold), and oocyte genes were more expressed in the study group (P < 0.0001) (~2.5-3.11 fold). These results were also confirmed by immunocytochemistry. The amount of steroids was much higher in the study group (three times and five times in two weeks) (P < 0.0001).

Conclusions: Stem cells can be obtained from the FF of PCOS, and normal FF has a positive effect on the growth and maturation of oocyte-like cells in vitro.

Keywords: Follicular fluid; PCOS; oocyte retrieval; stem cell; zona pellucida.

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Conflict of interest statement

There are no conflicts of interest.

Figures

Figure 1
Figure 1
Morphology of the cultured cells of the follicular fluid of PCOS (a) cells on the first day, (b) spindle-shaped cells on the second day, (c) fibroblast-like cells (F) and oocyte-like cells (O) in the third and fourth days, (d) epithelial-like cells (E) in the third and fourth days, (e) oocyte-like cells (O) in the control group in two weeks, and (f) oocyte-like cells (O) in the study group (with normal follicular fluid) in two weeks. (20X)
Figure 2
Figure 2
Expression of NANOG and OCT4 genes in stem cells and ZP2 and ZP3 in oocyte cells in the control group (with FBS) and study group (with follicular fluid) in the first and second weeks of culture. ****: P <0.0001, ***: P =0.009, ns = not significant
Figure 3
Figure 3
Immunofluorescence staining of stem cell markers (NANOG and OCT4) and oocyte-like cells (ZP2 and ZP3)
Figure 4
Figure 4
Determining the amount of estradiol in culture media containing normal follicular fluid (FFN) and without follicular fluid (FBS) in the first and second weeks ****: P <0.0001
See this image and copyright information in PMC

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