Neural stem cell differentiation in collagen scaffolds for retinal tissue engineering

Abstract

Rat neural stem cells (NSCs) were cultured in monolayer or in porous collagen scaffolds and exposed to neurogenic or non-neurogenic medium to determine the effects on neural differentiation and neurite growth. Nestin, [beta]III-tubulin, and GFAP expression were determined using immunofluorescent techniques, and the neurite length was measured. NSCs differentiated into neurons, with actively growing neurites, and astrocytes when cultured in differentiation medium (DM) or neurogenic medium (NM). NSCs cultured in monolayer expressed more nestin and III-tubulin and had significantly longer neurite extensions than NSCs cultured in collagen scaffolds. Laminin coated scaffolds promoted the attachment of NSCs to the scaffold struts and resulted in a more even distribution of nestin and [beta]III-tubulin positive cells throughout the scaffold. Overall, NSCs cultured in DM for at least 14 days resulted in the most neuronal differentiation and neurite growth.