Predictive Biophysical Cue Mapping for Direct Cell Reprogramming Using Combinatorial Nanoarrays

Biophysical cues, such as nanotopographies of extracellular matrix (ECM), are key cell regulators for direct cell reprogramming. Therefore, high-throughput methods capable of systematically screening a wide range of biophysical cueregulated cell reprogramming are increasingly needed for tissue engineering and regenerative medicine. Here, we report the development of a dynamic laser interference lithography (DIL) to generate large-scale combinatorial biophysical cue (CBC) arrays with diverse micro/nanostructures at higher complexities than most current arrays. Using CBC arrays, a high-throughput cell mapping method is further demonstrated for the systematic investigation of biophysical cue-mediated direct cell reprogramming. This CBC array-based high-throughput cell screening approach facilitates the rapid identification of unconventional hierarchical nanopatterns that induce the direct reprogramming of human fibroblasts into neurons through epigenetic modulation mechanisms. In this way, we successfully demonstrate DIL for generating highly complex CBC arrays and establish CBC array-based cell screening as a valuable strategy for systematically investigating the role of biophysical cues in cell reprogramming.

Automated summary

This study reports the development of a dynamic laser interference lithography (DIL) technique to generate large-scale combinatorial biophysical cue (CBC) arrays with diverse micro/nanostructures for investigating the role of biophysical cues in direct cell reprogramming. Through this approach, unconventional nanopatterns that induce the direct reprogramming of human fibroblasts into neurons were rapidly identified.