Colonization of Mouse Spermatogonial Cells in Modified Soft Agar Culture System Utilizing Nanofibrous Scaffold: A New Approach

  • Ali Talebi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Ali Sadighi Gilani Department of Urology, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran
  • Morteza Koruji Cellular and Molecular Research Center & Department of Anatomical Sciences, Iran University of Medical Sciences, Tehran, Iran
  • Jafar Ai Department of Tissue Engineering, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
  • Mohammad Jafar Rezaie Department of Embryology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
  • Shadan Navid Department of Anatomy, School of Medicine, Qom University of Medical Sciences, Qom, Iran
  • Majid Salehi Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
  • Mehdi Abbasi Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Keywords: Adult Germline Stem Cells, Cell Proliferation, Tissue Scaffolds, Agar


Background: Spermatogonial stem cells (SSCs) are considered in fertility management approaches of prepubertal boys facing cancer therapies. However, in vitro propagation has become an important issue due to a small number of SSCs in testicular tissue. The present study aimed to investigate a modified soft agar culture system by using a nanofibrous scaffold as a new approach to mimic in vivo conditions of SSCs development. Materials and Methods: The SSCs were isolated from neonate mouse testes, cultured on polycaprolactone scaffold, and covered by a layer of soft agar for 2 weeks. Then, the number and diameter of colonies formed in experimental groups were measured and spermatogonial markers (i.e., Plzf, Gfrα1, Id4, and c-Kit) in SSCs colonies were evaluated by a real-time polymerase chain reaction and immunostaining. Results: Our results indicated that the colonization rate of SSCs was significantly higher in the present modified soft agar culture system (P<0.05). Only Plzf indicated a significant increased at the levels (P<0.05), the gene expression levels of Id4, Plzf, and Gfrα1 were higher in the present culture system. In addition, the expression of the c-Kit gene as a differentiating spermatogonia marker was higher in presence of scaffold and soft agar compared with the amount of other experimental groups (P<0.05). Conclusion: The culture system by using nanofibrous scaffold and soft agar as a new culture method suggests the potential of this approach in SSCs enrichment and differentiation strategies for male infertility treatments, as well as in vitro spermatogenesis. [GMJ.2019;8:e1319]


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How to Cite
Talebi, A., Sadighi Gilani, M. A., Koruji, M., Ai, J., Rezaie, M. J., Navid, S., Salehi, M., & Abbasi, M. (2019). Colonization of Mouse Spermatogonial Cells in Modified Soft Agar Culture System Utilizing Nanofibrous Scaffold: A New Approach. Galen Medical Journal, 8, e1319.
Original Article