Z Narimanpour - Department of Anatomy & Cell Biology, Immunogenetic Research Center, Faculty of Medicine, Mazandaran University of Medical Sci-ences, Sari, Iran
M Nazm Bojnordi - Department of Anatomy & Cell Biology, Immunogenetic Research Center, Faculty of Medicine, Mazandaran University of Medical Sci-ences, Sari, Iran
S Ebrahimi - Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
J Saremi - Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran

Background: Nano fibrous scaffolds improve in vitro prolif-eration of stem cells via serving a similar and appropriate Ex-tra Cellular Matrix (ECM), guiding the cell attachment. In this study the stimulatory effects of a Silk nanofibrous scaffold was evaluated on maintain and in vitro proliferation rate of mouse spermatogonial stem cells (SSCs).Materials and Methods: Mouse neonatal spermatogonia stem cells were isolated from mouse testis (2-4days) using a modi-fied two-step mechanical and enzymatic digestion. After seed-ing SSCs in the sterilized silk scaffolds, they were incubated in DMEM/F12 medium. The attachment potential of SSCs were evaluated using SEM at days 3 and 5th days post seeding on the scaffold. MTT assessments and DAPI staining was used for viability and attachment of seed cells. Specific spermatogonia markers includes; Stra8, DAZL and Piwil2 were investigated via real time PCR and immunocytochemistry techniques.Results: MTT dates showed that cell viability rate of seeded SSCs on silk scaffold showed a significant increase in compari-son with the control group. The viability rate of SSCs post two weeks culture was 76.90% ± 4.16% and showed a significant increase (87.40% ± 0.73%) in cultured SSCs on silk scaffold. The attachment and proliferation of SSCs was increased in the presence of Silk electrospun scaffold in compare to monolayer sells. Real time PCR dates proved an upregulation in expres-sion of specific spermatogonial markers e.g.; Stra8, DAZL and Piwil2 in SSCs grown on silk scaffold, in compare with control group.Conclusion: It is concluded that Silk nanofibrous scaffold improves maintain and viability of neonatal mouse spermato-gonial stem cells, and stimulates the proliferation rate during cultivation. The results of our study suggests fabrication of the electrospun silk scaffold as a biological substitutes for efficient propagation of spermatogonial stem cells which is a crucial step in tissue engineering for reproductive medicine.

Spermatogonia Stem Cells, Scaffold,Silk, Prolifera-tion, Tissue Engineering