10.1021/am404134u.s012 Jong-Cheol Choi Jong-Cheol Choi Hong-Ryul Jung Hong-Ryul Jung Junsang Doh Junsang Doh Dynamic Modulation of Small-Sized Multicellular Clusters Using a Cell-Friendly Photoresist American Chemical Society 2013 multicellular cluster sizes cell micropatterning method multistep microscope projection photolithography multicellular clusters 2013-12-11 00:00:00 Media https://acs.figshare.com/articles/media/Dynamic_Modulation_of_Small_Sized_Multicellular_Clusters_Using_a_Cell_Friendly_Photoresist/2343760 Dynamics of small-sized multicellular clusters is important for many biological processes including embryonic development and cancer metastasis. Previous methods to fabricate multicellular clusters depended on stochastic adhesion and proliferation of cells on defined areas of cell-adhering islands. This made precise control over the number of cells within multicellular clusters impossible. Variation in numbers may have minimal effects on the behavior of multicellular clusters composed of tens of cells but would have profound effects on groups with fewer than ten cells. Herein, we report a new dynamic cell micropatterning method using a cell-friendly photoresist film by multistep microscope projection photolithography. We first fabricated single cell arrays of partially spread cells. Then, by merging neighboring cells, we successfully fabricated multicellular clusters with precisely controlled number, composition, and geometry. Using this method, we generated multicellular clusters of Madin–Darby canine kidney cells with various numbers and initial geometries. Then, we systematically investigated the effect of multicellular cluster sizes and geometries on their motility behaviors. We found that the behavior of small-sized multicellular clusters was not sensitive to initial configurations but instead was determined by dynamic force balances among the cells. Initially, the multicellular clusters exhibited a rounded morphology and minimal translocation, probably due to contractility at the periphery of the clusters. For 2-cell and 4-cell clusters, single leaders emerged over time and entire groups aligned and comigrated as single supercells. Such coherent behavior did not occur in 8-cell clusters, indicating a critical group size led by a single leader may exist. The method developed in the study will be useful for the study of collective migration and multicellular dynamics.