This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This is a pilot project of the Roadmap Consortium (U54 RR02437, T. Woodruff, PI) entitled "The Oncofertility Consortium: Fertility Preservation for Women." The process of folliculogenesis in vivo requires the coordinated development and maturation of the follicle wall and the oocyte enclosed within cumulus cells for eventual ovulation and fertilization. The ability to grow mouse follicles in a three-dimensional (3D) culture system yielding mature oocytes capable of fertilization and development to live offspring formed the basis of our current studies in nonhuman primates. We found that macaque follicles can survive, grow, form an antrum and secrete hormones when grown in encapsulated 3D culture. Despite exposure of macaque small antral follicles derived from encapsulated 3D culture to a maturation stimulus, most of the healthy oocytes remained immature. However, two oocytes matured to metaphase II, one fertilized via intracytoplasmic sperm injection and underwent early embryonic cleavage. Encapsulated 3D culture of macaque follicles to the small antral stage allows the functional gene expression of each follicular compartment (somatic cells, oocytes) to be analyzed separately and compared to follicles that develop to an equivalent stage/size in vivo. These studies will assess whether the encapsulated 3D system allows coordinated development of granulosa/theca cells and cumulus-oocyte complexes similar to that in vivo, and if not, will help define cellular functions that require further optimization in the 3D follicle culture system. This knowledge is critical for optimizing the 3D follicle culture technology for translation to human studies which will contribute to fertility preservation for female cancer survivors.