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

机译：用于用户指导的神经网络形成的可合成光降解水凝胶的设计和表征

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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 stem cell function and fate. With a growing interest in dynamically tunable cell culture systems, methods to synthesize photolabile hydrogels from simple 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 two-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.
机译：将光可裂解单元结合到交联中的水凝胶为研究人员提供了暂时控制机械性能并研究基质信号传导对干细胞功能和命运的作用的能力。随着人们对动态可调细胞培养系统的日益增长的兴趣，从简单的前体合成光不稳定水凝胶的方法将促进更广泛的可及性。在这里，开发了一种逐步生长的光降解聚乙二醇（PEG）水凝胶体系，该体系通过应变促进的炔-叠氮化物环加成（SPAAC）反应交联并通过整合到交联中的硝基苄基醚部分的裂解而降解。通过三个简单的合成步骤从商业上可获得的前体以高收率（> 95％）合成。响应于单光子和双光子辐照来表征网络的演化和降解特性。 PEG水凝胶用于封装胚胎干细胞衍生的运动神经元（ESMNs），并利用双光子红外光通过原位降解获得对运动轴突延伸的三维控制。最后，将ESMN及其体内突触伴侣肌管共囊化，并证明了用户控制的神经网络的形成。

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