The injury to the freeze-dried human spermatozoa is not only caused by the influence of preservation media to the physiological function, but also by the freezing and drying process. Now the selection of protective agents for the freeze-drying of sperm focuses on the view of physiological chemistry, aiming at restraining the enzyme activation in sperm. But the injury from freezing and drying has been omitted. Many studies have shown that egg yolk could alleviate the freezing injury, and disaccharides (especially for trehalose) could offer special protection to cells during drying. In this work, trehalose was added to the freeze-drying protective agents that were commonly used by others, or trehalose and egg yolk were added together. According to the experimental results of Eosin Y staining, hypoosmotic swelling test and observation by microscopy, the protective functions of these media to the structure and membrane of the sperm were compared. These results implies that the media modified by trehalose could improve the protection to sperm, and the media supplemented with trehalose and egg yolk is the best.
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