Spermatogonial stem cells (SCCs) is foundation for spermatogenesis throughout male adult life because they have ability of self-renewal and differentiation into spermatozoa. Storage of such SSCs is very important to study on male reproduction, which would contribute human male infertility to be treated. However, during cryopreservation, the most cells are damaged by cryoinjury such as apoptosis, necrosis, osmotic stress, oxidative stress and so on. For the reason, in cryopreservation technique, targeting purpose is what cells are stored stably without cryoinjury. The purpose of this study was to develop the cryoprotectant for decrease in cryoinjury of SSCs by using melatonin and necrostatin-1 as additive cryoprotectant. The SSCs with melatonin or necrostatin-1 was frozen for 1 month, and then thawed to evaluate survival, recovery and proliferation rate. The result showed that necrostatin-1 50 mM was significantly greater than DMSO control. Furthermore, we conducted the characterization of cryo-thawed SSCs with necrostatin-1 50 mM to confirm whether the SSCs could maintain the undifferentiated state. As a result, the normal expression of each marker, which is PLZF, GFRa1 and VASA, was observed except for C-kit, meaning that the cells could maintain the undifferentiated state regardless of cryopreservation. Therefore, the result indicates that the cryo-thawed SSCs have ability of proliferation and self-renewal. In conclusion, our finding verifies that cryopreservation of SSC with necrostatin-1 50 mM could be helpful to preserve the SSCs stably, contributing to various studies on male reproduction and infertility treatment

Lignocellulosic biomass conversion to biofuels such as ethanol and other value-added bio-products including activated carbons has attracted much attention. The development of an efficient, cost-effective, and eco-friendly pretreatment process is a major challenge in lignocellulosic biomass to biofuel conversion. Although several modern pretreatment technologies have been introduced, few promising technologies have been reported. Microwave irradiation or microwave-assisted methods (physical and chemical) for pretreatment (disintegration) of biomass have been gaining popularity over the last few years owing to their high heating efficiency, lower energy requirements, and easy operation. Acid and alkali pretreatments assisted by microwave heating meanwhile have been widely used for different types of lignocellulosic biomass conversion. Additional advantages of microwave-based pretreatments include faster treatment time, selective processing, instantaneous control, and acceleration of the reaction rate. The present review provides insights into the current research and advantages of using microwave-assisted pretreatment technologies for the conversion of lignocellulosic biomass to fermentable sugars in the process of cellulosic ethanol production.