Production of embryonic stem like-cells from neonatal mouse testis after exposure to low-intensity ultrasound

Pluripotent stem cells derived from testis are a new, unlimited source for cell therapy in regenerative medicine. Recently, studies show that spermatogonial stem cells can form embryonic stem-like cells (ES-like cells) in vitro. New procedures such as low intensity ultrasound (LIUS) can have positive effects on cell growth and differentiation. However, the effect of LIUS stimulation on ES-like cells has not been explored. In this study we investigate the effects of LIUS on colonization of ES-like cells.
Initially, we isolated SSCs from neonatal mice. The spermatogonial and Sertoli cells were cultured together in DMEM/F12 supplemented with 15% fetal bovine serum (FBS) and leukemia inhibitory factor (LIF). ES-like cells were stimulated by LIUS at intensity doses of 200 millwatt/square centimeter (mW/cm2) over 5 days. Characteristics of the isolated cells were confirmed by immunocytochemistry with Sox2 and SSEA-1 protein for ES-like cells. We also investigated colonization features in the ES-like cells.
After 21 days, we observed there was a significant increase in diameter and number of colonies in the 200 mW/cm2 group compared to the control group (p≤0.05). Pluripotency proteins, ES-like cell marker Sox2, and SSEA-1 expressed in the ES-like cells.
LIUS treatment can be an efficient, cost-effective method to improve colonization of ES-like cells during culture.