Design and Characterization of a Synthetically Accessible, Photodegradable Hydrogel for User-Directed Formation of Neural Networks

摘要

Hydrogels with photocleavable units incorporated into the cross-links have provided researchers with the ability to control mechanical properties temporally and study the role of matrix signaling on stenv cell fonction and fate. With a growing interest in dynamically tunable cell culture systems, methods to synthesize photolabile hydrogels front slrriple precursors would facilitate broader accessibility. Here, a step-growth photodegradable poly(ethylene glycol) (PEG) hydrogel system cross-linked through a strain promoted alkyne-azide cycloaddition (SPAAC)* reaction and degraded through the cleavage of a nitrobenzyl ether ;moiety integrated into the cross-links is developed from commercially available precursors in three straightforward synthetic Steps with high yields (>95%). The network evolution and degradation properties are characterized in response to one- and twd-photon irradiation. The PEG hydrogel is employed to encapsulate embryonic stem cell-derived motor neurons (ESMNs), and in situ degradation is exploited to gain three-dimensional control over the extension of motor axons using two-photon infrared light. Finally, ESMNs and their in vivo synaptic partners, myotubes, are coencapsulated, and the formation of user-directed neural networks is demonstrated.