No abstract

The aim of the present study was to evaluate the effect of activin A on the activation of in vitro folliculogenesis of human ovarian tissues from transgender men with or without our new compartmented chitosan hydrogel microbioreactor (“three-dimensional (3D)-structure”) enabling a three-dimensional tissue culture. Five fresh ovarian human tissues were cultured in vitro for 20 or 22 days in four groups with 100 ng/mL activin A or without activin A during the last six to eight days of culture, and within a 3D-structure or without the 3D-structure in standard conditions. Follicular density and quality were evaluated, and follicular diameters were measured. Estradiol secretion was quantified. Proliferation and apoptosis through immunostaining were also performed. The proportion of primordial follicles was significantly reduced, and the proportion of primary and secondary follicles was significantly increased in all four groups (p < 0.001). Tertiary follicles were observed in the four culture groups. Activin A supplementation did not significantly affect the follicular density, follicular quality, follicular growth, or estradiol secretion (p > 0.05). The 3D-structure increased the density of primary follicles and decreased the estradiol secretion (p < 0.001). Follicular proliferation was significantly lower in the 3D-structure group compared to the non-3D-structure group (p = 0.008). Regarding follicular apoptosis, it was significantly higher in the activin group compared to the non-activin group (p = 0.006). Activin A did not seem to play a key role in the in vitro folliculogenesis activation in our culture conditions. However, the results may indicate that the 3D-structure could be more physiological and could prevent a detrimental in vitro folliculogenesis flare-up.

Background:Cryopreservation of ovarian tissue is one of the strategies offered to girls and women needing gonadotoxic treatment, as a means of preserving their fertility. There are two methods of ovarian tissue cryopreservation: slow freezing, the reference method, and vitri cation, an alternative method. The aim of the present study was to evaluate which of the two is the best method for human ovarian tissue cryopreservation. Each ovary was divided into 3 groups: Fresh, Slow freezing and Vitri cation. In each group a histological study to evaluate follicular density and quality; and an evaluation of 6 gene expression (CYP11A, STAR, GDF9, ZP3, CDK2, CDKN1A) were performed. Results:We observed no signi cant difference in follicular density within these 3 groups. Slow freezing altered the pool of primordial follicles compared to the Fresh tissue (31.8% vs 55.9%, p = 0.046, respectively). The expression of genes involved in steroidogenesis varied after cryopreservation compared to the fresh group; CYP11A was under-expressed in both freezing groups compared to the fresh group and signi cantly under-expressed in the slow freezing group (p = 0.01), STAR was over-expressed in the slow freezing group and signi cantly under-expressed in the vitri cation group (p = 0.01). Regarding the expression of genes involved in cell cycle regulation, CDKN1A was signi cantly under-expressed in both freezing groups (slow freezing: p = 0.0008; vitri cation: p = 0.03) compared to the fresh group. Conclusion:Vitri cation had no effect on the histological quality of the follicles at any stage of development compared to Fresh tissue. There was no signi cant difference in gene expression between the two techniques.