Differentiation of Spermatogonial Stem Cells by Soft Agar Three-Dimensional Culture System Publisher Pubmed



Mohammadzadeh E^{1, 2} ; Mirzapour T³ ; Nowroozi MR⁴ ; Nazarian H¹ ; Piryaei A^{1, 5} ; Alipour F² ; Modarres Mousavi SM² ; Ghaffari Novin M¹ Show Affiliations
Source: Artificial Cells# Nanomedicine and Biotechnology Published:2019

Abstract

Since proliferation and differentiation of spermatogonial stem cells (SSCs) in culture system provide successful transplantation in this study, culture of human SSCs was compared to SACS (soft agar culture system), gelatin and control groups. The cells were isolated from seminiferous tubules of non-azoospermia patients (NOA) and cultured in DMEM for 3 weeks. The presence of SSCs in culture system was confirmed by immunocytochemistry of GFR-α1 and ITGα6 antibodies. The proliferated cells were cultured in three mentioned groups in the presence of retinoic acid and Sertoli cells conditioned medium for another 2 weeks. The number of colonies in the SACS group was significantly higher than two other groups. Before 2 weeks of culture, only Oct4 expression was observed in testicular cells (2.32 ± 0.25). After 2 weeks, the expression of Oct4 in the gelatin group was higher than that of the SACS group on day 7. The maximum expression of Stra8 was observed in SACS and gelatin groups after 7 days, but its expression was significantly decreased after 14 days of culture (p <.05). The expression of Scp3 and Acrosin genes were higher after 14 days in the SACS group compared to other groups. SACS has positive effects on proliferation and differentiation of hSSCs. © 2019, © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.