“…Isolated secondary follicles (~200 µm) improved their growth rates in the presence of increasing concentrations of FSH (100 ng/ml D0-D6; 500 ng/ml D6-D12; 1000 ng/ml D12-D18) alone (Saraiva et al, 2011) or in combination with LH and/or EGF (Saraiva et al, 2010;Silva et al, 2013). Moreover, sequential increasing FSH concentrations stimulated oocyte meiotic resumption to the GVBD stage when added alone (Saraiva et al, 2011) or in combination with IGF-I (Magalhães-Padilha et al, 2012b), while a low number of oocytes were able to reach the MII stage when sequential FSH was combined with: insulin (Chaves et al, 2012), IGF-II (Duarte et al, 2013), EGF (Silva et al, 2013), LH and EGF (Saraiva et al, 2010), GH (Magalhães et al, 2011), and VEGF (Araújo et al, 2011;Silva et al, 2014). Moreover, a few of these MII oocytes were fertilized and continued further development into 8-cell embryo (Silva et al, 2014), 16-cell embryo (Saraiva et al, 2010), and morula (Magalhães et al, 2011), which is the latest embryo developmental stage attained so far.…”