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Regulation of Mouse Follicle Development by Follicle-Stimulating Hormone in a Three-Dimensional In Vitro Culture System Is Dependent on Follicle Stage and Dose¹

Department of Chemical and Biological Engineering,³ Center for Reproductive Research,⁴ Department of Neurobiology and Physiology,⁵ Northwestern University, Evanston, Illinois 60208 Department of Medicine,⁶ Northwestern University, Chicago, Illinois 60611

The developmental requirements of ovarian follicles are dependent on the maturation stage of the follicle; in particular, elegant studies with genetic models have indicated that FSH is required for antral, but not preantral, follicle growth and maturation. To elucidate further the role of FSH and other regulatory molecules in preantral follicle development, in vitro culture systems are needed. We employed a biomaterials-based approach to follicle culture, in which follicles were encapsulated within matrices that were tailored to the specific developmental needs of the follicle. This three-dimensional system was used to examine the impact of increasing doses of FSH on follicle development for two-layered secondary (100–130 µm; two layers of granulosa cells surrounding the oocyte) and multilayered secondary (150–180 µm, several layers of granulosa cells surrounding the oocyte) follicles isolated from mice. Two-layered secondary follicles were FSH responsive when cultured in alginate-collagen I matrices, exhibiting FSH dose-dependent increases in follicle growth, lactate production, and steroid secretion. Multilayered secondary follicles were FSH dependent, with follicle survival, growth, steroid secretion, metabolism, and oocyte maturation all regulated by FSH. However, doses greater than 25 mIU/ml of FSH negatively impacted multilayered secondary follicle development (reduced follicle survival). The present results indicate that the hormonal and environmental needs of the follicular complex change during the maturation process. The culture system can be adapted to each stage of development, which will be especially critical for translation to human follicles that have a longer developmental period.

follicle, follicle-stimulating hormone, follicular development, oocyte development, ovary

² Correspondence: Lonnie D. Shea, Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Rd., Tech E136, Evanston, IL 60208. FAX: 847 491 3728; l-shea{at}northwestern.edu

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