Morphological and Molecular Aspects of In Vitro Culture of
Preantral Follicles Derived from Vitrified Ovarian
Tissues Using A Two-Step Culture
This study aimed to evaluate the expression of the genes related to folliculo-genesis after vitrification of mouse ovarian tissues using a two-step
Materials and Methods
In this experimental study, vitrified and non-vitrified ovaries from
7- day old (neonate) female mice were cultured using alpha-Minimum Essential Medium
(α-MEM) supplemented with 5% fetal bovine serum (FBS) for 7 days. Morphology, surface
area of ovaries and percentage of normal follicles were evaluated and compared in both
groups. After one-week culture, in non-vitrified group, preantral follicles of cultured ovaries
were isolated and cultured in a three-dimensional alginate culture system for 12 days.
Then, the collected metaphase (M) II oocytes were inseminated with capacitated spermatozoa derived from 7-8-week old (adult) male NMRI mice. Follicular diameter, oocyte maturation, fertilization, embryo development and the expression of genes related to follicular
The ovarian area in vitrified group (162468.20 703.78) was less than non-vitrified group (297211.40 6671.71), while the percentage of preantral follicles in vitrified group (18.40%) was significantly lower than those of non-vitrified group (24.50%) on day 7 of culture (P<0.05). There were no significant differences between the two groups in terms of follicular diameter, expression of genes related to development of follicles, oocyte maturation, fertilization, as well as embryo development (P>0.05).
The results of this study showed that vitrification of ovarian tissue following