Establishment of oxidative stress model during spermatogonial stem cells cultivation treated with different doses of H2O2

Barati, S.; Movahedin, M.; Mazaheri, Z.; Batooli, H.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p31
Academic Journal
Introduction: Nowadays, spermatogonial stem cells (SSCs) cultivation has been used as an effective tool for infertility treatments, by many researchers. Oxidative conditions can be effectiveness on cell proliferation and differentiation of these cells. So, the aim of this study was establishment of oxidative stress model for antioxidant activity of some drugs investigation during SSCs in vitro culture. Materials and Methods: Neonatal NMRI male mice (3-5 day) were used for isolation of SSCs. The cell suspension was prepared by twice enzymatic digestion. The cell suspension contents were spermatogonial and Sertoli cells and treated by different doses of H2O2 logarithmic concentrations from 0-1000 μM after 24 hours. To access the optimal dose, extra doses from 10-100 μM was evaluated. After 2 hours of H2O2 treatment, viability was determined by MTT assay. The data was analyzed using SPSS software and One-way ANOVA test. Results: Our data showed that spermatogonial cells colonies appeared after 4 days of isolation. These cells expressed OCT4 and PLZF proteins. Many of spermatogonial cells were removed after using of higher doses of H2O2. The results showed that 50 μM concentration of H2O2 could induce oxidative stress in spermatogonial stem cell during in vitro culture. Conclusion: According to this study, 50 μM concentration of H2O2 can cause cell death lower than 50% of total number of cells and increase oxidative stress in cultivation of SSCs. This model is a suitable tool for studying of some new antioxidant drugs.


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