Chitosan-g-oligo(L,L-lactide) Copolymer Hydrogel Potential for Neural Stem Cell Differentiation

We evaluated the applicability of chitosan-g-oligo(L,L-lactide) copolymer (CLC) hydrogel for central nervous system (CNS) tissue engineering. The biomechanical properties of the CLC hydrogel were characterized and its biocompatibility was assessing with neural progenitor cells obtained from two different sources: H9-derived neural stem cells (H9D-NSC) and directly reprogrammed neural precursor cells (drNPC). Our study found that the optically transparent CLC hydrogel possessed biomechanical characteristics suitable for culturing human neural stem/precursor cells and was non-cytotoxic. When plated on films prepared from CLC copolymer hydrogel, both H9D-NSC and drNPC adhered well, expanded and exhibited signs of spontaneous differentiation. While H9D-NSC mainly preserved multipotency as shown by a high proportion of Nestin+ and Sox2+ cells and a comparatively lower expression of the neuronal markers βIII-tubulin and MAP2, drNPCs, obtained by direct reprogramming, differentiated more extensively along the neuronal lineage. Our study indicates that the CLC hydrogel may be considered as a substrate for tissue-engineered constructs, applicable for therapy of neurodegenerative diseases.