Inhibiting EV Secretion Impacts NSC Differentiation

To investigate the effect of inhibiting the secretion of UTX KO SCMECs-derived EVs (KO EVs) or NC SCMECs-derived EVs (NC EVs) on NSCs differentiation, we need to co-culture SCMECs and NSCs. This study used a 24-well plate with 0.4 μm transwell chamber (Corning, 3413) to establish a co-culture system for SCMECs and NSCs. The SCMECs were seeded in the upper chamber of the transwell. In order to inhibit the secretion of SCMECs EVs, the upper complete medium of the experimental group contained 0.01% GW4869 (Sigma, D1692, GW4869 group), while the control group did not contain GW4869. GW4869 is an inhibitor of sphingomyelinase which can inhibit the biogenesis and release of EVs [37 (link)]. NSCs were digested into single cells and seeded in the lower chamber to construct the SCMECs-NSCs co-culture system.