Assessment of Survival and Expansion of Mouse Pre-Antral Follicles in 3D System Within Hyaluronic Acid-Alginate Hydrogel Cocultured With Human Theca Stem and Progenitor Cells Publisher Pubmed



Badban L ; Pirjani R ; Valojerdi MR
Source: Molecular Reproduction and Development Published:2026

Abstract

Cancer treatments can impair fertility in young women, and existing fertility preservation methods are often unsuitable for prepubertal patients. Replicating the natural follicular microenvironment in a three-dimensional (3D) in vitro culture of ovarian follicles presents a promising alternative, and co-culturing follicles with ovarian cells may enhance oocyte development. This study used a 3D culture system of hyaluronic acid and alginate to grow mouse preantral follicles and human theca stem and progenitor cells (hTSCs and hTPCs). Ovarian tissue was obtained from five transsexual women; hTSCs were isolated, expanded, confirmed, and differentiated into hTPCs. Preantral mouse follicles were collected and co-cultured with hTSCs and hTPCs in 0.5% hyaluronic acid-alginate (HA-Alg) hydrogel for 14 days. hTSCs were identified by spindle-shaped fibroblast morphology, positive alkaline phosphatase activity, expressing mesenchymal markers like CD29, CD44, and CD90, and differentiation potential to adipocytes and osteocytes-like cells. hTSCs differentiation into hTPCs causes an increase in steroidogenic gene expression, hormone release, and follicular recruitment. Coculturing of mouse preantral follicles with hTSCs and hTPCs in HA-Alg hydrogel induced follicle survival, hormone release, and oocyte maturation. Human theca cells in the stem and progenitor cells stage may support mouse follicle growth, survival, and steroidogenesis. © 2026 Wiley Periodicals LLC.