Synthesis of a Reversible Crosslinked Hydrogel

Abstract

Detachment of fragile cell types cultured on two-dimensional (2D) surfaces detrimentally affectscell viability. Accutase treatment of neurons growing in 2D damaged fragile neuronalconnections, resulting in cell death and host tissue damage in vivo. Avoiding cell detachmentaltogether by developing an in vitro culture system may better maintain cell viability; therefore,we present the synthesis of a reversible crosslinked three-dimensional (3D) hydrogel whichwould provide stability for culture/differentiation when crosslinked and injectability fortransplantation following crosslink degradation. A blend of hyaluronan (HA) andmethylcellulose (MC), HAMC, was chemically crosslinked with an enzyme-degradable peptidesequence using oxime click-chemistry. Specifically, HA and MC were modified with ketone andaldehyde groups, respectively, and a TEV protease-degradable peptide crosslinker wassynthesized and modified with oxyamine groups. The gel components were characterized, andgels were formed that demonstrated good stability, improved injectability upon crosslinkdegradation, and cytocompatibility with iPSC-derived neural progenitor cells