Evaluation of N-acetylcysteine (NAC) Effect on In Vitro Culture of Immature Mouse Testis Following Vitrifi-cation

Publish Year: 1398
نوع سند: مقاله کنفرانسی
زبان: English
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RROYAN20_327

تاریخ نمایه سازی: 29 مهر 1398

Abstract:

Background: Cryopreservation of testicular tissue in order to preserving spermatogonial stem cells (SSCs) is a suitable meth-od can be provided to prepubertal boys at the risk of infertility due to cancer treatments. Cryopreserved tissue can be trans-planted to the individual or cultivated in the laboratory. Due to the culture time, tissue damage is high, so optimizing the culture condition is essential. In this study, the effects of NAC presence in the culture medium of vitrified mouse testicular tis-sue will be investigated and cell viability will be evaluated.Materials and Methods: Testis tissue were harvested from sacrificed immature NMRI male mice and vitrified (freezing medium: DMSO, Ethylene Glycol, DMEM, 20% FBS) After warming, testis tissues were cultured in vitro for 7 days on agar gel in a medium composed of RPMI, NaHCO3 and 10% KSOR. Culture medium were supplemented by different dos-ages of NAC (0, 5, 10, 20, 25, 37.5, 50 mmol/L). Following 7 days of culture, cell viability were evaluated by Flow cytometry method.Results: Our results showed that cell survival At the end of the culture period was 54% at a NAC concentration of 50 mmol/L, 43% at 37.5 mmol/L, 39% at 25 mmol/L, 33% at 20 mmol/L, 22% at 10 mmol/L and 29% at 5 mmol/L. Cell survival was significantly higher in a group cultured without NAC compared to the groups cultured in presence of NAC (P<0.005).Conclusion: Although, our results showed that the used con-centrations of NAC wasn’t effective enough to suppress cell death in in vitro culture of vitrified testes, Previous researches established the ability of NAC to inhibit apoptosis in testicular germ cells in vitro. Thus, it seems that higher concentrations of this antioxidant should be tested.

Authors

P Nikoosokhan Tayar

Department of Developmental Biology, Faculty of Basic Scienc-es and Advanced Technologies in Biology, University of Science and Culture, Tehran, Iran. Department of Embryology, Reproductive Biomedicine Re-search, Royan Institute for Reproductive Biomedici

B Ebrahimi

Department of Embryology, Reproductive Biomedicine Re-search, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

A Alizadeh

Department of Embryology, Reproductive Biomedicine Re-search, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran

S Hajiaghalou

Department of Embryology, Reproductive Biomedicine Re-search, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran