Effects of Sperm Cryopreservation in Presence ofDeep Eutectic Solvents on Sperm Fertility Potential andROMO۱ Gene Expression

Background: The use of cryopreservation agents (CPAs) playsan important role in cell survival, as they prevent formation ofice crys tals. Deep eutectic solvents (DES) can act as CPA. Reactiveoxygen species (ROS) modulator (ROMO۱) interferes with the production of ROS. The reduction in the level of ROMO۱gene expression causes reduction in the ROS rate within cellsand improves sperm features. Therefore, this s tudy evaluatesthe effect of sperm cryopreservation in the presence of a specifictype of DES including choline chloride and glucose (ChG)on the expression of ROMO۱ gene and sperm quality.Materials and Methods: Normozoospermia samples were collectedfrom Mehr IVF center (Rasht, Iran). The semen sampleswere grouped as following: I. control (non-freeze), II. freezedwith SpermFreezeTM (Fertipro Co.), and III. freezed with DES(ChG). After thawing process, the samples were evaluated forsperm quality, including assessment of chromatin integrity (toluidineblue), chromatin condensation (aniline blue), survivaland acrosome reaction (triple s taining), and evaluation of spermmembrane integrity (eosin-nigrosin s taining). The expressionof ROMO۱ gene was detected by reverse transcription-quantitativePCR.Results: The results show that there is not a significant differencein the sperm quality (such as chromatin integrity and condensation,and intact acrosome) and sperm parameters (suchas viability, motility, morphology, councentration) followingfreezeing with ChG solvent compared to group II (P>۰.۰.۵).Also, ROMO۱ gene expression did not change in the ChGgroup compared to groups of I and II (P>۰.۰۵).Conclusion: The results of s tudy indicate that ChG is able to beconsidered as CPA and use for sperm cryopresvetion routinely.

کلیدواژه ها:

Deep Eutectic Solvents ، Male Infertility ، ROMO۱ ، Sperm Cryopreservation