Isolation, identification and differentiation of human spermatogonial cells on three-dimensional decellularized sheep testis

Ashouri Movassagh, Sepideh and Ashouri Movassagh, Sanaz and Banitalebi Dehkordi, Mehdi and Pourmand, Gholamreza and Gholami, Keykavos and Talebi, Ali and Esfandyari, Sahar and Jabari, Ayob and Samadian, Azam and Abbasi, Mehdi (2020) Isolation, identification and differentiation of human spermatogonial cells on three-dimensional decellularized sheep testis. Acta Histochemica, 122 (8). p. 151623. ISSN 00651281

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Abstract

Improvement of in vitro culture methods of Spermatogonial Stem Cells (SSCs) is known to be an effective procedure for further study of the process of spermatogenesis and can offer effective therapeutic modality for male infertility. Tissue decellularization by providing natural 3D and extracellular matrix (ECM) conditions for cell growth can be an alternative procedure to enhance in vitro culture conditions. In the present study, the testicular tissues were taken from brain death donors. After enzymatic digestion, the tissue cells were isolated and cultured for four weeks. Then the identity of the SSCs was confirmed using anti-GFRα1 and anti-PLZF antibodies via immunocytochemistry (ICC). The differentiation capacity of SSCs were evaluated by culture of them on a layer of decellularized testicular matrix (DTM) prepared from sheep testis, as well as under two-dimensional (2D) culture with differentiation medium. After four and six weeks of the initiation of differentiation culture, the pre-meiotic, meiotic and post- meiotic genes at the mRNA and protein levels was examined via qPCR and ICC methods, respectively. The results showed that pre-meiotic, meiotic and post-meiotic genes expressions were significantly higher in the cells cultured in DTM substrate (P ≤ 0.01).The present study indicated that, the natural structure of ECM prepare the suitable conditions for further study of the spermatogenesis process in the in vitro and contributes to the maintenance and treatment of male infertility.

Item Type: Article
Subjects: WJ Urogenital System
QU Biochemistry > Cell biology and genetics
Divisions: Reserach Vice-Chancellar Department > Cellular and Molecular Research Center
Depositing User: zeynab . bagheri
Date Deposited: 10 Nov 2020 10:19
Last Modified: 10 Nov 2020 10:38
URI: http://eprints.skums.ac.ir/id/eprint/8752

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