Micro-/nano-patterns on hydrogels are widely used in cell patterning.However,manufacturing molds with traditional lithog-raphy is time-consuming and expensive.In addition,the excessive demolding force can easily damage patterns since biocompatible hydrogels are ultra-soft or brittle.Here,we presented a novel method for rapid and mass fabrication of cell patterns.High-precision three-dimensional(3D)printing was used to manufacture a mold with a resolution of 2 μm,and the gelatin-based hydrogel was cured by thermal-photo-crosslinking so that the low-concentration and low-substitution-rate hydrogel could be demolded successfully.We found that pre-cooling before illumination made gelatin-based hydrogels resilient due to the partial regain of triple-helix structures.With this method,arbitrarily customized hydrogel patterns with a feature size of 6-80 μm can be fabricated stably and at low cost.When cardiomyocytes were seeded on ultra-soft hydrogels with parallel groove structures,a consistent and spontaneous beating with 216 beats per minute(BPM)could be observed,approaching the natural beating rate of rat hearts(300 BPM).Overall,this work provides a general scheme for manufacturing cell patterns which has great potential for cell ethology and tissue repair.

Chaofan He;Xuechun Chen;Yuan Sun;Mingjun Xie;Kang Yu;Jing He;Jinwei Lu;Qing Gao;Jing Nie;Yi Wang;Yong He

State Key Laboratory of Fluid Power and Mechatronic Systems,School of Mechanical Engineering,Zhejiang University,Hangzhou 310027,China;Pharmaceutical Informatics Institute,College of Pharmaceutical Sciences,Zhejiang University,Hangzhou 310058,China;Department of Orthopedic Surgery,The Second Affiliated Hospital,School of Medicine,Zhejiang University,Hangzhou 310009,China;State Key Laboratory of Component-Based Chinese Medicine,Tianjin University of Traditional Chinese Medicine,Tianjin 300072,China;Key Laboratory of Materials Processing and Mold,Zhengzhou University,Zhengzhou 450002,China;Key Laboratory of 3D Printing Process and Equipment of Zhejiang Province,College of Mechanical Engineering,Zhejiang University,Hangzhou 310027,China;Cancer Center,Zhejiang University,Hangzhou 310058,China

生物设计与制造（英文）

[1]Chaofan He;Xuechun Chen;Yuan Sun;Mingjun Xie;Kang Yu;Jing He;Jinwei Lu;Qing Gao;Jing Nie;Yi Wang;Yong He-.Rapid and mass manufacturing of soft hydrogel microstructures for cell patterns assisted by 3D printing)[J].生物设计与制造（英文）,2022(04):641-659

lithog,demolding,demolded,ethology

Rapid,mass,manufacturing,soft,microstructures,cell,patterns,assisted,by,printing,Micro,nano,hydrogels,are,widely,used,patterning,However,molds,traditional,raphy,consuming,expensive,In,addition,excessive,force,can,easily,damage,since,biocompatible,ultra,brittle,Here,presented,novel,method,rapid,fabrication,High,precision,three,dimensional,was,manufacture,resolution,gelatin,cured,thermal,photo,crosslinking,that,low,concentration,substitution,rate,could,successfully,We,found,cooling,before,illumination,made,resilient,due,partial,regain,triple,helix,With,this,arbitrarily,customized,feature,size,fabricated,stably,cost,When,cardiomyocytes,were,seeded,parallel,groove,consistent,spontaneous,beating,beats,per,minute,BPM,observed,approaching,natural,hearts,Overall,work,provides,general,scheme,which,has,great,potential,tissue,repair

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