Increasing of post-freezing quality of Spermatogonial Stem Cells after pretreatment by vitamin E
Mouse spermatogonial stem cells (SSCs) can be cryopreserved for long periods while preserving their spermatogenic ability. Although cryopreservation has been found to increase re- active oxygen species (ROS) formation that damages cellular structures. In the present study, we added vitamin E to the basic freezing medium in order to evaluate its effect on the efficiency of spermatogonial stem cells.
SSCs isolated from testes of 6 days old male mice by enzymatic digestion. Vitamin E 100, 200, 400 μg/mL was added to the basic freezing medium. The cell viability was evaluated by MTT assay. After thawing, SSCs were cultured for 1 month and the expression pattern of specific genes of SSCs measured by real-time PCR technique.
The survival rate of the freeze cells in the presence of vitamin E was significantly higher than the control group (p<0.05). The number of colonies and their diameter measured after one month were significantly higher in the vitamin E groups than in the control group (p≤0.05).
Adding vitamin E to the basic freezing medium thus can be helpful in increasing the quality and viability of SSCs after cryopreservation.
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