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Bio-Design and Manufacturing  2022 Vol.5 No.4 P.641-659

http://doi.org/10.1007/s42242-022-00207-1


Rapid and mass manufacturing of soft hydrogel microstructures for cell patterns assisted by 3D printing


Author(s):  Chaofan He, Xuechun Chen, Yuan Sun, Mingjun Xie, Kang Yu, Jing He, Jinwei Lu, Qing Gao, Jing Nie, Yi Wang & Yong He

Affiliation(s):  State Key Laboratory of Fluid Power and Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, Hangzhou 310027, China; more

Corresponding email(s):   zjuwangyi@zju.edu.cn, yongqin@zju.edu.cn

Key Words:  3D printing, Gelatin-based hydrogel, Cell patternCardiomyocytes beating


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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]. Journal of Zhejiang University Science D, 2022, 5(4): 641-659.

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Abstract: 
Micro-/nano-patterns on hydrogels are widely used in cell patterning. However, manufacturing molds with traditional lithography 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.

【封面文章】浙大贺永、王毅等 | 生物水凝胶微结构快速制造方法助力心肌细胞定向可控生长

本研究论文聚焦助力心肌细胞定向可控生长的生物水凝胶微结构快速制造方法。生物水凝胶上的微/纳米结构广泛用于诱导细胞图案化。传统的光刻技术制造模具既耗时又昂贵,并且由于生物水凝胶软脆的特性,过大的脱模力极易导致微结构损坏。本文提出一种快速批量制造细胞图案的新方法,使用高精度3D打印模具,通过揭示热-光交联固化机制,实现低浓度-低取代率的GelMA水凝胶微结构高质高效制造(最低为7%浓度-30%取代率)。利用该方法,可以稳定且低成本地制造特征尺寸为6–80 μm任意定制的水凝胶图案。在超软水凝胶及高精度微结构的支持下,心肌细胞可实现每分钟216次(BPM)的持续自发搏动,接近大鼠心脏的自然搏动速率(300 BPM)。这项工作为细胞图案化提供了一个通用方案,能广泛应用于组织修复、药物测试筛选等领域。

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