Hierarchical Patterning of Cells with a Microeraser and Electrospun Nanofibers

For tissue engineering applications, it is important to develop fabrication strategies for building models with controlled cell distributions in defined structures. Here, a simple, flexible approach (named the μ-eraser strategy) is developed to construct multicell micropatterns. This approach involves pressing a poly(dimethylsiloxane) stamp to erase cells growing on substrates, and seeding other types of cells. The pressing/seeding process can be conducted in any designed pattern at desired time point. In a proof of concept, multicell micropatterns of human lung adenocarcinoma epithelial A549 cells, murine fibroblast (FB) cells and murine osteoblast (OB) cells are achieved on Petri dishes and electrospun sheets. Besides forming multicell micropatterns, the cell orientation can be regulated by microstripes and alignment of nanofibers. On Petri dishes and random fiber sheets, FB and OB cells align along microstripes, while A549 cells do not. However, when growing on aligned fiber sheets, no matter whether solo-cultured or co-cultured, all cells in micropatterns orient along the fibers. Based on this technique, a platform is built up to investigate rates of cell migration and interinvasion under solo-culture and co-culture systems. It is believed that this μ-eraser strategy has promise for biological, pharmaceutical, and biomedical applications.