The mitochondrial DNA copy number, cytochrome c oxidase activity and reactive oxygen species level in metaphase II oocytes obtained from in vitro culture of cryopreserved ovarian tissue in comparison with in vivo-obtained oocyte.

Aim: To evaluate the mitochondrial DNA (mtDNA) copy number, reactive oxygen species (ROS) level and intensity of mitochondrial enzyme activity in metaphase II oocytes derived from vitrified cultured immature mouse ovarian tissue in comparison with nonvitrified group and in vivo-obtained oocytes.

Methods: Vitrified and nonvitrified ovaries from neonate female mice were cultured for 7 days. Then, preantral follicles were isolated and cultured in a three-dimensional culture system. Follicular development and oocyte maturation were evaluated and compared in both groups. Some of the collected metaphase II oocytes derived from in vitro and in vivo conditions were inseminated with capacitated spermatozoa, and then, the fertilization and embryo developmental rates were assessed. In the other series of oocytes, mtDNA copy number, distribution and enzyme activity and ROS level were analyzed.

Results: The embryo development, mtDNA copy number and mitochondrial enzyme activity in collected metaphase II oocytes from two in vitro-cultured groups were significantly lower, and the ROS level was higher than those of the in vivo group (P < 0.05), but there was no significant difference between vitrified and nonvitrified groups.

Conclusion: This study showed that a two-step in vitro culture of mouse ovarian tissue decreased the mtDNA copy number and cytochrome c oxidase activity of metaphase II oocytes through an increase in their ROS level in comparison with in vivo-obtained oocytes. Thus, the in vitro culture methods should be improved.