In the present study, we evaluated the contributions of antral follicle development and antral granulosa cell-released factor(s) to the acquisition of a mature mouse oocyte plasma membrane organization and cortical granule distribution. This has been performed by comparing in-vitro matured oocytes derived from early antral follicles (here referred to as denuded oocytes) or from pre-ovulatory follicles, and cultured either as cumulus-intact or cumulus-free oocytes, with in-vivo ovulated eggs. By using scanning and transmission electron microscopy, the denuded oocyte surface appears to be characterized by the presence of long microvilli, while that of pre-ovulatory oocytes and of ovulated eggs by shorter microvilli. However, denuded oocytes can acquire a pre-ovulatory-like plasma membrane configuration when matured in vitro in the presence of early antral granulosa or cumulus cells, but not of NIH-3T3 fibroblasts. On the contrary, fluorescence and confocal microscopy analyses after labelling with fluorescent Lens culinaris agglutinin show that all the oocyte classes analysed are characterized by similar cortical granule distribution and density. Thus, complete antral follicle development plays an important role in the process of oocyte surface differentiation, probably through the action of antral granulosa cell-released factor(s), but it does not affect oocyte capacity to normally distribute cortical granules.