BACKGROUND: Cryopreservation is a technique by which, semen can be preserved to subzero temperature, usually at -196° C. The freezing of semen desires vitrification mediators that diminish wreck to the cells (spermatozoan) during the freeze and thaw process. Using cryopreservation, the quality of the semen has been increased in the latest years, by which the achievement rate for the insemination techniques has increased in an agreed way. The area need to be focused is to enhance the quality of the semen extender preparation before cryopreservation. Many researchers are working in the area of cryopreservation of human semen with different semen extenders. Several parameters influence the properties of semen extender essential for better post thaw results. This review is mainly focused on a range of parameters which influence the best semen extender for cryopreservation that includes glycerol and its importance, buffer and novel usage of antimicrobial peptides as antimicrobial agents.