Survival and hormone production of isolated mouse follicles in three-dimensional artificial scaffolds after stimulation with bpV(HOpic)

Abstract

Abstract Purpose: To preserve fertility before gonadotoxic therapy, ovarian tissue can be removed, cryopreserved, and transplanted back again after treatment. An alternative is the artificial ovary, in which the ovarian follicles are extracted from the tissue, which reduces the risk of reimplantation of poten-tially remaining malignant cells. The PTEN inhibitor bpV(HOpic) has been shown to activate human and bovine ovarian follicles, and it is therefore considered a promising substance for de-veloping the artificial ovary. Methods: A comparative analysis was performed, studying four different groups that differed either in the scaffold (polycaprolactone scaffold versus polyethylene terephthalate membrane) or in the medium — bpV(HOpic) versus control medium. The observation period was 10 days. On days 2, 6, and 10, the viability and morphology of the mice follicles were checked using fluorescence or confocal microscopy. Furthermore, hormone levels of estrogen (pmol/L) and progesterone (nmol/L) were determined. Results: When comparing the survival rates (SR) of follicles among the four groups, it was observed that on day 6, the study groups utilizing the polycaprolactone scaffold with bpV(HOpic) in the me-dium (SR: 0.48 ± 0.18) or functionalized in the scaffold (SR: 0.50 ± 0.20) exhibited significantly higher survival rates compared to the group using only the polyethylene terephthalate membrane (SR: 0.34 ± 0.15). On day 10, a significantly higher survival rate was only noted when comparing the polycaprolactone scaffold with bpV(HOpic) in the medium to the polyethylene terephthalate membrane group (SR: 0.38 ± 0.20 versus 0.21 ± 0.11). Higher levels of progesterone were partly associated with better survival rates, but only reached significance in the group with the poly-caprolactone scaffold functionalized with bpV(HOpic). Conclusion: This study demonstrates that three-dimensional polycaprolactone scaffolds improve the survival rates of isolated mice follicles in comparison with a conventional PET membrane. Higher rates of progesterone were also partly associated with improved survival.