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Adv Pharm Bull. 2021;11(4): 712-718.
doi: 10.34172/apb.2021.080
  Abstract View: 366
  PDF Download: 27

Research Article

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

Soudabe Yousefi 1,2,3 ORCID logo, Maryam Akbarzadeh 4 ORCID logo, Jafar Soleimanirad 1, Kobra Hamdi 5, Laya Farzadi 5, Aalie Ghasemzadeh 5, Mahdi Mahdipour 1, Reza Rahbarghazi 1,5* ORCID logo, Mohammad Nouri 1,5* ORCID logo

1 Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
2 Department of Biochemistry and Clinical Laboratories, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
4 Stem Cell and Regenerative Medicine Institute, Tabriz University of Medical Sciences, Tabriz, Iran.
5 Department of Applied Cell Sciences, Faculty of Advanced Medical Sciences, Tabriz University of Medical Sciences, Tabriz, Iran.
6 Women’s Reproductive Health Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
*Corresponding Author: Mohammad Nouri, Tel:+98 41 33373879; Fax: +98 41 33363870, Email: Nourimd@yahoo.com

Abstract

Purpose: Previous studies have documented that cumulus granulosa cells (GCs) 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 GCs toward the oocyte-like phenotype in vitro.

Methods: GCs were isolated from healthy female volunteers subjected to in vitro fertilization or intra-cytoplasmic sperm injection (IVF-ICSI). The effect of different media supplemented with leukemia inhibitory factors (LIFs), 5 ng/mL estradiol, and 0.005 IU/mL follicle-stimulating hormone (FSH) were investigated to the differentiation of GCs 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 polymerase chain reaction (PCR) assay. We also assess morphological adaptation by using bright-field microscopic imaging.

Results: Exposure of GCs to LIFs 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 FSH (P<0.05). According to our data, the incubation of GCs with estradiol increased the expression of genes related to the oocyte-like phenotype.

Conclusion: Our finding revealed that the combination of LIFs and estradiol could induce the GCs’ oogenesis capacity and thereby is possibly suggested as a therapeutic strategy during the occurrence of gynecological disorders.


Keywords: Granulosa cells, Leukemia inhibitory factors, Follicle-stimulating hormone, 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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