Effect of Mycotoxins on spermatogonial stem cells

Abstract

Fusarium mycotoxins are natural contaminants of various commodities representing significant problem worldwide. Since beauvericin (BEA), one of the fusarium toxins, represents a major concern because of its potential toxicity in humans and animal health, and its high presence in feed and food commodities, the present study was carried out to evaluate the effect of BEA on cell viability, proliferation, morphology and gene expression using spermatogonial stem cells (SSCs). First, SSCs were isolated from fresh sheep testis under aseptic technique. Isolated SSCs were cultured for 36 hours at 5% CO2. Then, they were treated with different chosen BEA dose for different time points. Cell count using the hemocytometer was done to determine cell viability. BEA at 0.3, 1, and 3μM significantly decreased (p <0.05) cell numbers after 12 and 24 hours of treatment. For the percentage of viable cells MTT test was performed. At concentration ≥0.3μM, BEA was found to decrease (p <0.05) percentage of cell viability. Culture of SSCs on a slide treated with 1 and 3 μM for 24 hours shows a morphological change after the hematoxylin and eosin staining: elongated and dispersed cells with no more clustered SSCs. RNA extraction and amplification using PCR, then the expression of Oct-4, and Smad8/9 were assessed using qRT-PCR.BEA had no effect on Oct-4 and Smad8/9 mRNA abundance (p >0.05). Higher expression of Oct-4 (p <0.05) was found in SSCs indicating proliferation. Whereas, a lower expression of smad8/9(p >0.05) was shown supporting the purity of SSCs isolated. Taken together these results demonstrate that BEA may impair reproductive function in sheep.