Abstract
BackgroundThe mechanism of theca cell layer formation in mammalian ovaries has not been elucidated; one reason is that there is no follicle culture system that can reproduce theca cell layer formation in vitro. Therefore, a three-dimensional follicle culture system that can reproduce theca cell layer formation is required.MethodsA collagen gel was used in the follicle culture system. To determine the optimum conditions for follicle culture that can reproduce theca cell layer formation, the effects of hormonal treatment and cell types co-cultured with follicles were examined. In addition, immunohistochemistry was used to examine the properties of the cell layers formed in the outermost part of follicles.ResultsFollicles maintained a three-dimensional shape and grew in collagen gel. By adding follicle-stimulating hormone (FSH) and co-culturing with interstitial cells, the follicles grew well, and cell layers were formed in the outermost part of follicles. Immunohistochemistry confirmed that the cells forming the outermost layers of the follicles were theca cells.ConclusionIn this study, follicle culture system that can reproduce theca cell layer formation in vitro was established. In our opinion, this system is suitable for the analysis of theca cell layer formation and contributes to our understanding of the mechanisms of folliculogenesis.
Highlights
The mechanism of theca cell layer formation in mammalian ovaries has not been elucidated; one reason is that there is no follicle culture system that can reproduce theca cell layer formation in vitro
Three-dimensional follicle culture system Effects of collagen gel culture on follicular growth The follicles used in this culture system were obtained from 10- to 14-day-old mice and were approximately 140 μm in diameter
Effect of follicle-stimulating hormone (FSH) on follicular growth Because it has been suggested that FSH may play a role in follicular growth, we examined the effects of FSH on follicular development in collagen gels for 5 days
Summary
The mechanism of theca cell layer formation in mammalian ovaries has not been elucidated; one reason is that there is no follicle culture system that can reproduce theca cell layer formation in vitro. A three-dimensional follicle culture system that can reproduce theca cell layer formation is required. The follicles in mammalian ovaries are composed of a single oocyte, granulosa cells, and theca cells. The theca cell layer is not recognizable in primordial follicles, it is identifiable from the stage of secondary follicles onward. It is considered that theca cells play roles in the physical maintenance of follicle structure by constructing cell layers around the basement membrane. Knowledge of theca cells, theca cell layer formation, is less than that of granulosa cells and oocytes. The origin of theca cells has not been fully elucidated.
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