Study On Mouse Spermatozoa Transported At Ambient Temperature And Preserved At Refrigerated Temperatures

Objective:To explore the optimal conditions for cauda epididymides transport under ambient temperature and eliminate the effect of transport medium on the transport of cauda epididymides. To investigate the feasibility of cauda epididymides spermatozoa maintained in a refrigerator at4℃. To establish a system for valuable male mouse unexpected die and endangered strains conservation through assisted reproductive technology. Methods:The transport methods of cauda epididymides were modified in this study and the sperm motilities, fertility and developmental abilities of IVF embryos after transportation at room temperatures or4～8℃in20μl of transport medium or transport medium-free tube were compared. Cauda epididymides of B6D2F1、ICR and C57BL/6mice were stored in a refrigerator at4℃. The effects of storage time for sperm motilities, fertility and developmental abilities of IVF and ICSI embryos were explore. Results:The results showed that the transport medium-free delivery of cauda epididymides under room temperatures was significantly superior to the room temperatures delivery of cauda epididymides with transport medium, but was inferior to the4～8℃delivery of cauda epididymides in20μl transport medium. With the increase of storage time, the sperm motilities and fertility were significantly reduced. Within4days B6D2F1got the highest perm motilities and fertility, ICR took the second place and C57BL/6followed.45.5%fertilization rate obtained using cauda epididymides spermatozoa stored at4℃for60days when injected the sperm into oocytes. Conclusion:Thus the modified transport medium-free system was beneficial to room temperature transportation of cauda epididymides and could eliminate the side effects of transport medium on spermatozoa, and the delivery of cauda epididymides under room temperature was useful for transporting some valuable strains between mouse banks and research facilities. The4℃medium-free storage of cauda epididymides could get considerable fertilization rate of IVF and ICSI. This method allowed for easy long-term storage of mice spermatozoa with very low cost and wide application. In conclusion. this procedure could be particularly important in the unexpected die mice and endangered species.