en بيولوژي Biology پژوهشي Research <div style="text-align: justify;"><strong>Background</strong>: This study aimed to explore whether the addition of a cyclic adenosine monophosphate (cAMP) analog and isobutylmethylxanthine (IBMX) in freezing media improved sperm quality and what role cAMP has in this recovery.<br> <strong>Materials and methods:</strong> ach semen sample was cryopreserved into four groups: fresh semen sample, as a control group, freezing medium + 2.5 mM cAMP analog and 0.2 mM IBMX, freezing medium + 12.5 mM cAMP analog and 0.2 mM IBMX, and freezing medium + 25 mM cAMP analog and 0.2 mM IBMX. Sperm parameters after post-thaw were analyzed according to WHO instruction (2010). Viability, acrosome reaction, and DNA damage levels of the samples were evaluated.<br> <strong>Results</strong>: Our results indicated that the effective concentrations of 12.5 and 25 mM cAMP analog and 0.2 mM IBMX significantly improved the total motility, progressive motility, and viability of the frozen-thawed (P<0.05). However, non-progressive motility and immotile were significantly reduced in the 12.5 and 25 mM cAMP analogs and 0.2 mM IBMX groups after thawing (P<0.05). During freezing the spermatozoa, the high concentration of the cAMP analog increased acrosome reaction after thawing in the 25 mM and 0.2 mM IBMX treated samples (P<0.05). DNA fragmentation in 25 mM cAMP analog and 0.2 mM (IBMX) supplementation was significantly lower compared to the other groups (P<0.05).<br> <strong>Conclusions</strong>: Our findings revealed that in vitro cAMP analog and IBMX supplementation in freezing media play an important role in preventing cryodamage by maintaining the sperm functional parameters.</div> cAMP analog (IBMX), cryopreservation, Asthenozoospermic, sperm quality 201 208 http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1147-1&slc_lang=en&sid=1 Rahil Jannatifar rahiljannatifar2016@gmail.com 10031947532846009146 10031947532846009146 No Department of Reproductive Biology, the Academic Center for Education, Culture and Research, Qom branch, Iran Hamid Piroozmanesh hp457@yahoo.com 10031947532846009147 10031947532846009147 No Department of Reproductive Biology, the Academic Center for Education, Culture and Research, Qom branch, Iran Leila Naserpoor leilanasery48@gmail.com 10031947532846009148 10031947532846009148 Yes Department of Reproductive Biology, the Academic Center for Education, Culture and Research, Qom branch, Iran