Controlling cellular cytoskeletal arrangement, contractility, and uniformity is critical for precisely and uniformly regulating differentiation, gene expression, and motility for applications such as cell manufacturing, single cell analysis, and tissue engineering. Nanopatterned substrates are an ideal platform for manipulating single cells as this size regime is commensurate with the protein assemblies that anchor cells to the extracellular matrix and direct cytoskeletal element. However, the subcellular arrangement of cell adhesion ligands into arbitrary shapes remains challenging with conventional lithographic methods which limit our ability to precisely modulate biophysically derived cell behavior. To overcome this challenge, we use polymer pen lithography (PPL), a cantilever-free lithographic technique that utilizes millions of polymeric pyramidal pens, to pattern nanoscale features on substrates. Importantly, the maskless nature of PPL allows one to rapidly prototype unique nanopattern libraries utilizing the same pen array and thus facilitating high-throughput screening. Here, PPL is used to arrange cell adhesion proteins, such as fibronectin, into anisotropic features that direct the formation of highly oriented actin cytoskeletons within a defined shape factor. This controlled orientation results in a highly uniform and contractile cytoskeleton that allow one to enhance or alter the differentiation fate of human mesenchymal stem cells.

Subcellular control over focal adhesion anisotropy, independent of cell morphology, dictates stem cell fate

2019 Fall National Meeting

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