The effect of adding Rosmarinic and Ascorbic acids to vitrification media on fertilization rate of the mice oocyte: An experimental study|
Borjizadeh, Abdollah; Ahmadi, Hamid; Daneshi, Erfan; Roshani, Daem; Fathi, Fardin; Abdi, Mahdad; Nasseri, Sherko & Abouzaripour, Morteza
Background: Oocytes vitrification is a pivotal step for the widespread and safekeeping
of animal genetic resources. Oocytes endure notable morphological and functional
damage during cryopreservation. Oxidative stress is one of the adverse effects that
vitrification imparts on oocytes.
Objective: In the present study, we investigated the antioxidant effect of Rosmarinic
and Ascorbic acids on the quality and fertilizing ability of frozen-thawed mice oocyte.
Materials and Methods: In this experimental study, germinal vesicle oocytes obtained
from two-months-old (30–40gr) NMRI mice were randomly divided into four groups.
The basic cryoprotectants were 7.5% (v/v) ethylene glycol+7.5% (v/v) Propanediol as
an equilibration media. Vitrification medium contained 15% (v/v) ethylene glycol+15%
(v/v) propanediol, and 0.5 M sucrose. In the first group (Control), nothing was added
to vitrification mediums, whereas, in the second and third groups, 0.5 mmol/L of
Ascorbic acid and 105 μmol/L of Rosmarinic acid were added into vitrification medium,
respectively. The cumulative concentration of Rosmarinic and Ascorbic acids were
added to group 4. Mouse oocytes were vitrified and preserved for one month. The
thawed oocytes were transferred into the α-MEM medium (Alpha Minimum Essential
Medium) and maintained in this medium for 24 hr, to be matured and reach the
metaphase II stage.
Results: The addition of Rosmarinic and Ascorbic acids to the vitrification solution
improved the survival, maturation of Germinal vesicles, fertilization rate, and finally
development to 4-cell stage. Maturation rates to 4-cell stage for Ascorbic acid, Rosmarinic
acid, and both of them together were 80%, 80.76%, and 86.61%, respectively.
Conclusion: These results indicate that the addition of a cumulative concentration of
0.5 mmol/L Ascorbic acid and 105 μmol/L of Rosmarinic acid to the cryopreservation
solution for the mouse immature oocytes would be of significant value (p< 0.01).
Vitrification; Antioxidant; Fertilization.