Towards the Development of an Artificial Ovary

Ovarian preantral follicles often flatten and rupture during static culture in a tissue culture well. In this study, growth and morphology were evaluated for rat and mouse preantral follicles cultured for 72 hours in conventional culture wells and in suspension culture systems, consisting of orbiting test tubes and rotating-wall vessels. Follicles cultured in the orbiting test tubes had increased growth rates relative to the rate in the conventional culture wells, and experienced neither flattening nor rupture. The majority of follicles cultured in the rotating-wall vessel appeared to experience shear damage. In order to provide a potential barrier from shear stress during suspension culture, follicles were encapsulated in calcium alginate gels and growth and morphology were evaluated in the conventional and suspension culture systems. Encapsulated follicles more closely resembled an in vivo morphology and did not flatten nor rupture. Our studies suggest that suspension culturing in a rotating-wall vessel in combination with microencapsulation supports more natural three-dimensional follicular growth and morphology. The approach described herein is the first step toward the development of an artificial ovary. Additionally, this new culture system maintains follicles in a more natural morphology and will provide an important new avenue for further detailed investigation of the complex regulation of ovarian follicle development