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. 2018 Mar 1;8:3849. doi: 10.1038/s41598-018-21989-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Various patterns at the single-cell level in collagen produced with the drop-patterning method. (a) Photograph of the drop-patterned collagen gel in a 35-mm glass-bottomed Petri dish with millimeter-scale tumor patterns growing for six days from the single-cell level. (Scale bar: 15 mm) (b) Fluorescence images (Hoechst, maximum projection) of cell-assembled capital letters R, P, and I (abbreviation for Rensselaer Polytechnic Institute) after six days of growth in the collagen. (c) Single cells and small cell clusters were drop-patterned in collagen in concentric circles demonstrating the capability of producing varying intercellular spacing within one pattern (double-headed arrows). Ninety-nine of a total of 128 positions were occupied by cells (78% patterning efficiency). (d) Six-day culture of the cell pattern in (c) showing MCTS growth, cell migration, and cell protrusions in between MCTS (red and black solid lines). The radius R of the green circle is 500 μm. Blue dotted ellipses indicate two separate cells in (c) proliferating and merging into one MCTS in (d) after six days. (Scale bars: 200 μm).