The Effect of Nanocomposite Scaffold on Spermato-gonial Stem Cells Proliferations

Objective: Some male survivors of childhood cancer are in distress from azoospermia. Spermatogonail stem cells (SSCs) isolation and purification are really important. Testicular orga-noids and three-dimensional (۳D) scaffolds are possible appli-cations in treatment of male infertility. ۳D nanofibers scaffolds play important role in cell culture, and due to these scaffolds provide a microenvironment similar to extracellular matrix for proliferation and self-renewal of cells. The present study aimed to evaluate the efficiency of ۳D microenvironment containing the chitosan-alginate –Graphene oxide nanocomposites for cell culture improvement and proliferation of spermatogonial cells. Materials and Methods: Spermatogonial cells were cultured and divided into ۳ culture groups: ۱. Control (culture in basic media), ۲. SSC culture in CA/GO۳۰µg scaffold, and ۳. SSC culture on alginate-chitosan. The identity of the cultured cells was confirmed by flowcytometry (ckit and GFRα۱).The Scaf-folds were analyzed using FTIR, XRD, and scanning electron microscopy (SEM) to observe surface topography and the mor-phology .Cytotoxicity of scaffolds was assayed at ۲۴ hours, ۷۲ hours and one week after seeding using MTT assay. The stem cells related markers for SSCs (Id۴, GFRα۱ and PLZF) were detected on all experimental groups by quantitative Real-Time PCR (qRT-PCR).Results: These results showed that SSCs can easily attach and proliferate on the substrates. The CA/GO۳۰µg scaffold was biocompatible, as evidenced by the MTT assay. SSC that were seeded onto the surface of the scaffold exhibited good prolif-eration. The quantity of proliferations marker significantly in-creased compared with control group. The qRT-PCR results confirmed that GO-based chitosan-alginate scaffold may pro-vide an ideal environment for SSC proliferations.Conclusion: We conclude that SSCs culture in the group CA/GO۳۰µg has a potential use for SSCs proliferation in vitro. This ۳D scaffold is applicable for culturing and encapsulation of SSCs.