Worthen Christal A, Rittié Laure, Fisher Gary J
Department of Dermatology, University of Michigan Medical School, Ann Arbor, MI, USA.
Dermatology Therapeutic Area, GlaxoSmithKline, Collegeville, PA, USA.
Methods Mol Biol. 2017;1627:245-251. doi: 10.1007/978-1-4939-7113-8_17.
Polyacrylamide hydrogels can be used to culture cells in a range of stiffness that can closer mimic physiological environments. Changes in environmental stiffness have been documented in conditions such as fibrosis, cancer, and aging. In this chapter, we describe a method in which we pour gels directly into multiwell plates using a plastic support that covalently binds to the polymerizing hydrogel. The hydrogel is then crosslinked to calfskin collagen using a crosslinker. The result is a thick hydrogel, scalable to any size plate, which covers the entire surface of the well with no edge effects. The gels can be routinely assembled and are easily reproducible. These scaffolds are used as in vitro models to study fibroblast reaction to variation in environmental stiffness.
聚丙烯酰胺水凝胶可用于在一系列能更接近模拟生理环境的硬度条件下培养细胞。在诸如纤维化、癌症和衰老等病症中，已记录到环境硬度的变化。在本章中，我们描述了一种方法，即使用与聚合水凝胶共价结合的塑料支撑物将凝胶直接倒入多孔板中。然后使用交联剂将水凝胶与小牛皮胶原蛋白交联。结果得到一种厚水凝胶，可扩展到任何尺寸的板，它覆盖孔的整个表面且无边缘效应。这些凝胶可常规组装且易于重现。这些支架用作体外模型来研究成纤维细胞对环境硬度变化的反应。
