Using natural honey as non-permeating cryoprotectant: An approach to reduce apoptose in vitrified-warmed mouse blastocyst

Background: Vitrification process optimization, as a way to store embryos derived from assisted reproductive techniques, requires dealing with several destructive factors that expose the embryos to extreme conditions and leads to various stresses such as oxidative stress, subsequently apoptosis. The presence of carbohydrates as non-permeating cryoprotectants is an attempt in order to reduce these stresses. Since the combination use of sugars leads to better results, in this project, natural honey which consists of various monosaccharides and disaccharides was applied as an alternative to sucrose in mouse blastocyst vitrification.Methods: For this purpose, the first part of the experiment was conducted to determine the optimum concentration of honey in vitrification and warming solutions. The second part was designed to compare the effect of honey-based selected solutions and sucrose-based one in terms of survival, hatching and implantation rate, as well as the expression level of Tp53 and Bax genes.Result: Among different concentrations of honey, optimum concentrations of 1 and 2 M were chosen for vitrification and thawing mediums, respectively. Results of the second part demonstrated that, not only the survival rate can be equal with sucrose when using honey, but it also brings about as good results as the sucrose-based group regarding the hatching rate and more importantly the implantation rate. In addition, the increased expression level of Tp53 and Bax genes in sucrose-based group is modified by using the honey and approaches the non-vitrified control group.Conclusion: Therefore, we can introduce natural honey, as an appropriate substitute for sucrose to optimization the vitrification process.

کلیدواژه ها:

Apoptose ، Mouse blastocyst ، Natural Honey ، Non-permeating Cryoprotectant ، Tp53 and Bax gene expression ، Vitrification