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  Abstract View: 77

Research Article

Combination of Estradiol with Leukemia Inhibitory Factor Stimulates Granulosa Cells Differentiation into Oocyte-Like Cells

Soudabe Yousefi ORCID logo, Maryam Akbarzadeh ORCID logo, Jafar Soleimanirad, Kobra Hamdi, Laya Farzadi, Aalie Ghasemzadeh, Mahdi Mahdipour, Reza Rahbarghazi* ORCID logo, Mohammad Nouri

Abstract

Purpose Previous studies have documented that cumulus granulosa cells can trans-differentiation into different non-ovarian cells, showing their multipotentiality to repopulate the injured cells in ovarian tissue. The current experiment is aimed to assess the differentiation capacity of human cumulus granulosa cells toward the oocyte-like phenotype in vitro. Methods Granulosa cells were isolated from healthy female volunteers subjected to in vitro fertilization or intra-cytoplasmic sperm injection. The effect of different media supplemented with leukemia inhibitory factors, 5 ng/ml estradiol, and 0.005 IU/ml follicle-stimulating hormone were investigated to the differentiation of granulosa cells toward oocyte-like phenotype via monitoring the expression of Oct3/4 and GATA-4 using flow cytometry analysis. The expression of genes such as FIGLA, NOBOX, and SYCP3 was measured by real-time PCR assay. We also assess morphological adaptation by using bright-field microscopic imaging. Results Exposure of granulosa cells to leukemia inhibitory factors increased the number of cells expressing stemness factor Oct3/4 coincided with the suppression of GATA-4 after 7 days (p<0.05). We found that the transcript level of all genes FIGLA, Nobox, and SYCP-3 decreased in cells after treatment with a follicle-stimulating hormone (p<0.05). According to our data, the incubation of granulosa cells with estradiol increased the expression of genes related to the oocyte-like phenotype. Conclusion Our finding revealed that the combination of leukemia inhibitory factors and estradiol could induce the granulosa cells' oogenesis capacity and thereby is possibly suggested as a therapeutic strategy during the occurrence of gynecological disorders.
Keywords: Granulosa cells, Leukemia inhibitory factors, FSH, Estradiol, Oocyte-like cells
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Submitted: 11 Feb 2020
Revision: 29 Apr 2020
Accepted: 26 Jul 2020
ePublished: 26 Jul 2020
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