Effect of Acoustic Cavitation on Mouse Spermatogonial Stem Cells: Colonization and Viability.

Objectives: The mechanical index has long been one of the main criteria used to assess the safety limits for therapeutic medical applications. However, the safety of the mechanical index parameter is considered to be unknown in male fertility, which has a very significant role in vitro conditions. In this study, the effect of cavitation interactions due to mechanical index regions was evaluated on spermatogonial stem cells.

Methods: The acoustic pressure and mechanical index equations at the low intensities and the intended frequency were modeled and solved. The mechanical index average of 40 kHz frequency was selected as subthreshold, 0.70, and above the cavitation threshold. Neonatal spermatogonial stem cells were cultured. Spermatogonial stem cells are stimulated by low-level ultrasound for 5 days and colonization and viability evaluated on the seventh day.

Results: Based on modeling, the mechanical index average was chosen as 0.40, 0.51, 0.75, and 0.89. The mechanical index of 0.40 and 0.89 resulted in a number of colonies of 93 ± 4 and 32 ± 4, respectively. An increase in colony diameter could be observed for a 0.40 mechanical index during all days of the culture that in the culture on the seventh day had the largest average colony diameter of 174.05 ± 1.22 μm in comparison with other groups (p < 0.05). The cell viability was not significantly different among the groups.

Conclusion: The results suggest that a low-intensity ultrasound of 40 kHz with a 0.40 mechanical index can be effective in increasing the proliferation and colonization of spermatogonia in stem cells during culture.