Synthesis and mechanical characterization of polyacrylamide (PAAm) hydrogels with different stiffnesses for large-batch cell culture applications

Charlotte Cresens, Samet Aytekin, Behrad Shaghaghi, Lotte Gerrits, Ana Montero-Calle, Rodrigo Barderas, Paul Kouwer, Susana Rocha
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Abstract

The impact of mechanical cues on cell behavior is increasingly being recognized, rendering hydrogel platforms that mimic the extracellular matrix indispensable in in vitro cell biology research. Here, we present a step-by-step protocol for synthesis and rheological characterization of polyacrylamide (PAAm) hydrogels with varying stiffnesses, produced as large circular unattached gels customizable in shape and size. We outline methods for their use in cell culture and downstream applications involving secretome or cell analysis, and protein visualization by fluorescence microscopy.
用于大批量细胞培养的具有不同硬度的聚丙烯酰胺(PAAm)水凝胶的合成与力学性能分析
人们越来越认识到机械线索对细胞行为的影响,因此在体外细胞生物学研究中,模拟细胞外基质的水凝胶平台不可或缺。在这里,我们将逐步介绍不同硬度的聚丙烯酰胺(PAAm)水凝胶的合成和流变学特性。我们概述了将其用于细胞培养和下游应用的方法,这些应用涉及分泌物组或细胞分析,以及通过荧光显微镜观察蛋白质。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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