In the present study, we employed the harvesting media and the PCM with the same temperature and osmotic pressure. Therefore, the deterioration of the viability and Metaraminol bitartrate proliferation of NSCs is not caused by the temperature or osmolarity of the harvesting media but likely associated with pH alteration and a lack of nutrients. The experimental harvesting media do not contain any growth factors and only provided short-term survival of NSCs. Deficiency or exhaustion of exogenous instructive factors or lack of pH buffer system may greatly inhibit the viability and proliferation of cells in vitro, while a greater number of NSCs survive in the media containing sufficient nutrients, powerful buffer system and essential growth factors. Although we did not establish a relationship between the viability or proliferation of NSCs and the individual harvesting media in this study, our results suggest that NSCs cannot be maintained in the long term in the experimental harvesting media and indicate that appropriate harvesting media and appropriate treatment durations have benefits on cell survival and proliferation. However, it is Cortisone acetate striking that neurospheres can be maintained in the long term in the harvesting media. The mechanism underlying the differences in viability and proliferation and the tolerance to harvesting media exposure between dissociated NSCs and neurospheres remains to be identified. There is abundant evidence for the coupling of proliferation and cell cycle progression to the nutrient environment and pH alteration. We demonstrated that the exposure to the experimental group harvesting media triggers p53-mediated cell cycle arrest and represses the expression of cyclin E1, eventually leading to the reduction of S-phase entry. However, our understanding of how the harvesting media exposure affects cell cycle progression is limited. Cell cycle arrest during starvation is often mediated by an accumulation of cyclin-dependent kinase inhibitors, such as p27 and p21. However, p21, which acts downstream of p53, was not detected in this study, most likely due to its absence in embryonic stem cells as reported previously.