Of note, NSCs were exposed to the harvesting media at 4uC where NSCs are isolated, harvested or transported. Although the internalization of EGF receptor which is considered as the main mitogen receptor is completely blocked at 4uC, the EGF stimulation-induced phosphorylation and activation of EGFR and its effectors as well as their interaction are preserved and even enhanced. These may act as the Tirofiban molecular basis of the EGF function on NSCs at 4uC, which conforms to the sustained proliferation of NSCs exposed to the PCM at 4uC. Collectively, our results suggest that NSC cell cycle progression is responsive to the nutrient content and pH of harvesting media. Previous research indicates that growth factors have a significant survival function against cellular death. The harvesting media employed in the current study do not contain any of the growth factors, thus deprivation of growth factors in the harvesting media may result in apoptosis and necrosis of NSCs. Our data indicate that harvesting media exposure-associated NSC death including apoptosis and necrosis occurs in a time-dependent manner. The rate of apoptosis and necrosis in harvesting media exposure groups is Saline.PBS.ACSF. With prolonged treatment, the viable population of cells gradually shifts to the necrotic and apoptotic cell death population in the harvesting media exposure groups, and the progression is irreversible. Apoptosis is often initiated by either extrinsic or intrinsic signaling pathways. To further understand the molecular mechanism underlying NSC apoptosis, we examined the levels of (-)-Huperzine A apoptosis-related molecules. Our results indicate that following prolonged treatment duration, apoptotic pathways are activated. The expression of Fas-L and cleaved caspase 8, in NSCs are up-regulated, while activated caspase 9 in NSCs is not detectable. Additionally, fluorescence labeling assays reveal the intact mitochondrial inner membrane. According to our findings, it seems that the extrinsic pathway is involved in harvesting media exposure-associated NSC apoptosis.