Nylon mesh-based 3D scaffolds for the adherent culture of neural stem/progenitor cells.

Abstract

We developed novel scaffolds for the adherent culture of neural stem/progenitor cells on the woven mesh. Nylon mesh (NM) is an inert material for cell adhesion. We prepared polyacrylic acid-grafted nylon mesh (PAA-NM) by graft polymerization method using gamma-irradiation. Matrigel was covalently immobilized to the carboxyl groups in PAA-NM by chemical conjugation using 1-ethyl-3-(3-dimethylamino propyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) to prepare the Matrigel-immobilized PAA-grafted nylon mesh (M-PAA-NM). Cell adhesion property of mouse neural stem/progenitor cells (NSPCs) between the NM, PAA-NM, and M-PAA-NM was different from each other. The neurosphere-like clusters of NSPCs were weakly bound to NM and PAA-NM without spreading. The NSPCs were firmly adhered to, spread, and covered the surface of M-PAA-NM. We evaluated the state of differentiation by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immnocytochemistry. A neuronal marker β III tubulin, a glial marker glial fibrillary acidic protein (GFAP) and a mature glial marker S100β were expressed at a low level in the cultured cells while immature NSPCs marker Nestin and Sox2 were slightly lower without significant statistical difference. We concluded that the M-PAA-NM is a good substrate for adherent culture of NSPCs without triggering their cell differentiation, and also provides the maintenance of their growth with fewer passages in comparison with the conventional suspension culture of NSPCs in neurospheres.