Title: Colonization of Mouse Spermatogonial Cells in Modifed Soft Agar Culture System Utilizing Nanofbrous Scaffold: A New Approach
Journal: Galen Medical Journal
Author: 1. Ali Talebi, Shadan Navid, Mehdi Abbasi, 2. Mohammad Ali Sadighi Gilani, 3. Morteza Koruji, 4. Jafar Ai, 5. Mohammad Jafar Rezaie, 6. Majid Salehi
Year: 2019
Address: 1. Department of Anatomy, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran 2. Department of Urology, Shariati Hospital, Tehran University of Medical Sciences, Tehran, Iran 3. Cellular and Molecular Research Center & Department of Anatomical Sciences, Iran University of Medical Sciences, Tehran, Iran 4. Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran 5. Department of Embryology, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran 6. Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
Abstract: 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 modifed soft agar culture system by using a nanofbrous 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 signifcantly higher in the present modifed soft agar culture system (P<0.05). Only Plzf indicated a signifcant 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 nanofbrous 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.
Keywords: Adult Germline Stem Cells, Cell Proliferation, Tissue Scaffolds, Agar
Application: Scaffold
Product Model 1: Syringe Pump
Product Model 2: High Voltage Power Supply
URL: #https://www.researchgate.net/profile/Ali_Talebi/publication/332972048_Colonization_of_Mouse_Spermatogonial_Cells_in_Modified_Soft_Agar_Culture_System_Utilizing_Nanofibrous_Scaffold_A_New_Approach/links/5cd458d4a6fdccc9dd9a4815/Colonization-of-Mouse-Spermatogonial-Cells-in-Modified-Soft-Agar-Culture-System-Utilizing-Nanofibrous-Scaffold-A-New-Approach.pdf#