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

Abstract

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.