Neural stem cell encapsulation and differentiation in strain promoted crosslinked polyethylene glycol-based hydrogels

Encapsulated cell viability within crosslinked hydrogels is a critical factor to consider in regenerative medicine/cell delivery applications. Herein, a "click" hydrogel system is presented encompassing 4-dibenzocyclooctynol functionalized polyethylene glycol, a four arm polyethylene glycol tetraazide crosslinker, tethered native protein attachment ligands (laminin), and a tethered potent neurogenic differentiation factor (interferon-γ). With this approach, hydrogel formation occurs via strain-promoted, metal-free, azide-alkyne cycloaddition in an aqueous buffer. This system demonstrated safe encapsulation of neural stem cells in biological conditions without chemical initiators/ultraviolet light, achieving high cell viability. Cell viability in click gels was nearly double that of ultraviolet exposed gels after 1 d as well as 14 d of subsequent culture; demonstrating the sensitivity of neural stem cells to ultraviolet light damage, as well as the need to develop safer encapsulation strategies. Finally, protein immobilized click hydrogel neural stem cell in vitro differentiation over 2 weeks demonstrated that the click gels specified primarily neurons without the need for additional protein differentiation factor media supplementation.