The effect of encapsulating the ovaries with alginate hydrogels on the expression of caspase3 gene in the vitrified ovaries of mice

Background: Cryopreservation of the ovarian tissue is one of the methods for preserving fertility, which, unlike cryopreservation of the oocyte and embryo, can be done before puberty and at each stage of the menstrual cycle. Apoptosis may be induced by the process of cryopreservation due to changes in physical conditions. When induced by apoptosis in the cell, the cell death cascade activates the caspases and, as a results of cell damage, can affect the quality of the ovarian reserve; Encapsulation is a method that has been investigated and proposed to protect tissues and cells in different conditions.Objective: Therefore, due to the importance of apoptosis after vitrification and warming, the aim of this study was to determine the effect of hydrogel alginate as capsule on the expression of caspase 3 gene in the vitrified mouse ovaries.Materials and Methods: In this experimental study, the ovaries of adult mice were divided into 4 groups: non-vitrified ovaries, vitrified ovaries, ovaries encapsulated in alginate hydrogel at 0.5 and 1% concentration and then vitrified (respectively, Experimental group 1 and 2). Vitrification was performed with ethylene glycol and dimethyl sulfoxide solutions. The duration of exposure to liquid nitrogen was 30 minutes and warming the ovary with decreasing sucrose solutions. The level of expression of caspase3 gene was evaluated in each group with real time RT-PCR. The HPRT gene was considered as the reference gene. Data were analyzed by one-way ANOVA and post hoc tests. The values of p<0.05 were considered statistically significant.Results: Molecular studies showed that caspase3 gene was expressed in all groups. Non-vitrified group was considered as 1 and the other groups were compared to it. The expression level of this gene was higher in the experimental groups than in the vitrified group, but no significant difference was found between the experimental and the vitrified group. The lowest expression of this gene was observed in the experimental group 2 (1% concentration).Conclusion: Encapsulation of the ovaries with concentrations of 0.5 and 1% of alginate hydrogels did not reduce the expression of caspase3 gene in vitrified ovaries.

کلیدواژه ها:

Vitrification ، Alginate hydrogel ، Apoptosis ، Ovary ، Mice