Quality of Oocytes Derived from Vitrified Ovarian Follicles Cultured in Two- and Three-Dimensional Culture System in the Presence and Absence of Kit Ligand

Abstract

The aim of this study was to evaluate the effects of Kit Ligand (KL) on the growth of vitrified follicle, oocyte quality, and embryo development in two- and three-dimensional culture systems. Vitrified and nonvitrified mouse whole ovaries were cultured for 1 week, then their isolated preantral follicles were cultured for 12 days in two- or three-dimensional culture systems in the presence and absence of KL. The growth and diameter of follicles, maturation of oocytes, and hormonal level were assessed. Finally, embryo developmental rate and oocytes reactive oxygen species (ROS) levels, adenosine triphosphate (ATP) content, and distribution of mitochondria were examined. The results showed the maturation and survival rates and steroidogenesis of follicles were significantly higher not only in the three-dimensional culture system but also in the presence of KL (p < 0.001). The embryo development, ATP content, and ROS level of collected oocytes were not significantly different in all groups. Mitochondrial distribution within the oocyte was similar in all groups of the study. Our results demonstrated that KL supplementation during follicular culture in two- and three-dimensional culture systems appeared to significantly increase follicular function and development. The ovarian vitrification had no harmful effect on the steroidogenesis, growth, and maturation of follicles.

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

“…Abdi et al . also demonstrated that the mouse ovarian vitrification had no harmful effects on the steroidogenesis, growth and maturation of isolated follicles after two‐step in vitro culture of ovarian tissue . In the other point of view, the fertilization rate of MII oocytes derived from vitrified ovaries was significantly lower as compared with MII oocytes derived from the in vivo group.…”

“…The isolated preantral follicles ( n > 150 in each group) from cultured ovaries were encapsulated in sodium alginate as described previously . Encapsulated follicles were cultured in α‐MEM medium supplemented with 5% FBS, 100 mIU/mL rFSH or Gonal‐f (Serono), 1% ITS (Gibco), 100 mg/mL penicillin and 50 ng/mL streptomycin under mineral oil at 37°C with 5% CO 2 for 12 days.…”

“…IVM could be a method for monitoring the effect of the vitrification procedure on follicular development and oocyte maturation. In spite of great improvements in the ovarian tissue vitrification method, there are challenges to the effect of this method on the development of follicles …”

Effects of Matricaria chamomilla Extract on Growth and Maturation of Isolated Mouse Ovarian Follicles in a Three-dimensional Culture System

“…Furthermore, every other day, half of the medium was replaced by fresh medium. [ 22 ] At the end of culture period, analysis of developmental genes ( PCNA and FSHR ) and steroid hormones (17-β estradiol, progesterone, and dehydroepiandrosterone (DHEA)) was performed and also the released metaphase II (MII) oocytes in three groups of culture were collected and randomly considered for reactive oxygen species (ROS) assay.…”

“…The preantral follicle was encapsulated in sodium alginate. [ 22 ] Briefly, alginate was dissolved in deionized water at a concentration of 1% (w/v) and was then mixed with activated charcoal (0.5 g of charcoal per gram of sodium alginate). Following charcoal treatment, the prepared sodium alginate solution was sterile filtered through 0.22-μM filters.…”