The main findings of this work were that the short-term WZ4002 removal of growth factors is capable of modifying the distribution of btubulin III and GFAP positive cells in whole neurospheres; increasing the number of b-tubulin III positive cells, while decreasing the number of Nestin positive cells inducing changes in gene expression profiles; and promoting neurite extension and changes in orientation of neural stem cells. Here we show the ability of hNPC to survive and differentiate after culturing in suspension in the absence of growth factors. It has recently been shown that mNPC are able to survive and differentiate after 14 days growth factors starvation. However, MFM neurospheres did not grow as much as the CTR ones, as observed in growth rates. As neurosphere size is established by the balance between proliferation and cell death, the stable size of MFM neurospheres is due to both proliferation and apoptosis decreasing. As expected, BrdU incorporation decreases after mitogens removal. On the other hand, we have to take in account that, even after growth factors removal, the level of apoptotic cells was extremely low , again, showing that neurospheres should have a mechanism that prevents cell death in this condition. The increased apoptosis after 14 days in culture in the CTR group can be attributed to the increase in neurosphere size. Up to now, studies aiming at finding EWSFLI1 target genes investigated exclusively binding to promoter regions either through genome wide approaches or through specific analyses of genes transcriptionally modulated by this oncogene. In order to identify EWS-FLI1 specific in vivo target genes in an unbiased genome wide approach, we used here chromatin immunoprecipitation coupled with high throughput sequencing. The former correspond to the binding sites that are expected for the EWS-FLI1 factor, considering its common binding properties with wild type FLI1. Our approach not only broadens the list of such sites as EWS-FLI1 direct targets, but also points out their significant association in pairs or with other transcription factors binding sites within modules. The association of ETS binding sites with binding sites for factors such as CREB or NFkB may suggest a cooperative interplay of EWS-FLI1 with other cancer-related factors. The present identification of GGAA microsatellites as EWS-FLI1 targets confirms and extends a previous ChIP-on-chip-based, genome-wide analysis of EWS-FLI1 binding sites in promoter regions. Indeed, GGAA microsatellites were recently described as EWS-FLI1 binding sites within different promoters, including NROB1, FCGRT and caveolin 1. Moreover, EWS-FLI1 direct interaction with these repeated elements was validated by gel shift assays. This hypothesis is based on human and animal studies linking early growth and nutrition to long term risk of age-related diseases such as type 2 diabetes. There is also limited data suggesting a link between early life events and the aging process in humans.