This chapter focuses on human follicle culture and briefly describes follicle culture in some of the animal models that were inspiring for progress in humans. Several culture media used to grow human follicles in vitro have been supplemented with the growth factors to assess their potential physiological significance in regulating the earliest stages of follicular development in human. The ovarian cortical tissue is mainly populated with primordial follicles. In vitro follicle culture involved the use of isolated early-stage follicles. There are two approaches to the isolation of follicles: mechanical (microdissection) and enzymatic (use of enzyme). Basically, three major approaches in follicle culture techniques can be distinguished: culture of ovarian cortical tissue (humans and larger animals), culture of entire ovaries (rodents), and culture of isolated follicles (humans, larger animals, and rodents). Several histomorphometric methods are used to investigate follicular development in vitro.