Аннотация:Abstract Objectives The main purpose of current study was to explore the effects of tetrahedral DNA nanostructures (TDNs) on neuroectodermal ( NE ‐4C) stem cells migration and unveil the potential mechanisms. Materials and methods The successfully self‐assembled TDN s were also determined by dynamic light scattering ( DLS ). A bidirectional wound‐healing assay and transwell chamber assay were employed to test the migrating behaviour of NE ‐4C stem cells cultured under different conditions. Results Through an in vitro study, we found that stem cells could internalize TDN s quickly, and the cells’ parallel and vertical migration was promoted effectively. Besides, the effects of TDN s were found being exerted by upregulating the gene and protein expression levels of RhoA , Rock2 and Vinculin , indicating that the RHOA / ROCK 2 pathway was activated by the TDN s during the cell migration. Conclusions In conclusion, TDN s could enter NSC s without the aid of other transfection reagents in large amounts, whereas only small amounts of ss DNA could enter the cells. TDN s taken up by NSC s activated the RHOA / ROCK 2 signalling pathway, which had effects on the relevant genes and proteins expression, eventually promoting the migration of NE ‐4C stem cells. These findings suggested that TDN s have great potential in application for the repair and regeneration of neural tissue.