Since the groundbreaking discovery of induced pluripotent stem cells (iPSCs) many research groups have attempted to improve the efficiency of the classical cell reprogramming process. Surprisingly, the contribution of the three-dimensional (3D) microenvironment to iPSC generation has been largely overlooked. Here we describe a protocol for the generation of iPSCs in defined poly(ethylene glycol) (PEG)-based hydrogels that, besides allowing higher reprogramming efficiency, are also a powerful tool to study the influence of biophysical parameters on iPSC generation.
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