Covalent bonding of GYIGSR to EVAL membrane surface to improve migration and adhesion of cultured neural stem/precursor cells.

In the present study, we modified poly (ethylene-co-vinyl alcohol) (EVAL) membranes with the covalent bonding of the laminin-derived peptides, GYIGSR by using carbodiimidazole (CDI) to activate the hydroxyl groups on the membrane surface. The resulting GYIGSR-immobilized EVAL (EVAL-GYIGSR) membrane was analyzed in terms of the effect of immobilized peptide sequence on the behaviors of neural stem/precursor cells (NSPCs), isolated from embryonic rat cerebral cortex, in the serum-free medium. Compared to the unmodified EVAL, GYIGSR immobilized on the EVAL membrane was shown to significantly increase NSPCs migrating out of neurospheres (p<0.05). In addition, NSPCs on the EVAL-GYIGSR membrane were able to differentiate into neural lineage cells and differentiated neurons expressed functional synaptic activity. Basically, there was no significant difference between GYIGSR-immobilized and laminin-coated substrates for their ability to enhance migration and differentiation of NSPCs, suggesting that the immobilization of GYIGSR on the EVAL membrane was successful and the laminin-derived peptide YIGSR and laminin had similar effect on NSPC behaviors. However, it is non-permanent modification for coating laminin on the substrates to support cell survival after a long-term culture. In this study, differentiated neurons could still adhere to the EVAL-GYIGSR surface with functional synaptic activity after incubation for 20 days. Therefore, the bioactive EVAL-GYIGSR provided an alternative approach to improve migration and survival of NSPCs for neural tissue engineering applications.