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Date of download: 7/11/2016 Copyright © ASME. All rights reserved. From: Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting J. Nanotechnol. Eng. Med. 2015;6(2):021006-021006-5. doi:10.1115/1.4031217 Viability of printed cells. One day after printing, D1 cells adhered to collagen (left panel). The Live/Dead assay suggested that cells were mainly alive (middle panel), any dead cell was detected (right panel). Figure Legend:
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Date of download: 7/11/2016 Copyright © ASME. All rights reserved. From: Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting J. Nanotechnol. Eng. Med. 2015;6(2):021006-021006-5. doi:10.1115/1.4031217 Postprinting DNA fragmentation. Four hours after printing, D1 cells were fixed and treated for a TUNEL assay. No fluorescent staining was detected suggesting that there was no DNA fragmentation induced by LAB (left panel). A positive control (nuclease- treated sample, right panel) for the assay is presented. Green fluorescence corresponded to DNA terminal ends. Figure Legend:
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Date of download: 7/11/2016 Copyright © ASME. All rights reserved. From: Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting J. Nanotechnol. Eng. Med. 2015;6(2):021006-021006-5. doi:10.1115/1.4031217 Comparison of printed and unprinted cell proliferation. First, an Alamar Blue assay was performed for different seeding densities. Thus, the fluorescent signal detected was quantified in function of cell density (cells/cm 2 ). Then, an Alamar Blue assay was performed over 4 days. Results for printed (white) and unprinted (black) cells are presented. Figure Legend:
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Date of download: 7/11/2016 Copyright © ASME. All rights reserved. From: Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting J. Nanotechnol. Eng. Med. 2015;6(2):021006-021006-5. doi:10.1115/1.4031217 Ability of cells to differentiate toward osteogenic lineage after the printing process. Printed pluripotent precursors of bone marrow were grown in control medium (control) or in osteoblastic-inductive medium (differentiated). The absorbance measures correspond to the quantity of calcium produced by the cells. Figure Legend:
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Date of download: 7/11/2016 Copyright © ASME. All rights reserved. From: Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting J. Nanotechnol. Eng. Med. 2015;6(2):021006-021006-5. doi:10.1115/1.4031217 LAB of the cell patterns with high definition. (a) D1T cells, stained by tdTomato, were printed on a collagen layer. The position of each islet of cells varied from 150 μm (top panel) to 500 μm (bottom panel) between two successive islets within a line, with a 50 μm step between each panel. (b) The distance measured between each pair of consecutive islets on the substrate (d m ) was calculated using a dedicated python computer program and plotted in function of the distance expected (d e ). Figure Legend:
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Date of download: 7/11/2016 Copyright © ASME. All rights reserved. From: Creation of Highly Defined Mesenchymal Stem Cell Patterns in Three Dimensions by Laser-Assisted Bioprinting J. Nanotechnol. Eng. Med. 2015;6(2):021006-021006-5. doi:10.1115/1.4031217 High-definition cell printing in 3D. A first pattern of D1T cells was printed on a collagen layer (left panel). This pattern was covered with a second collagen layer. By changing the microscope focus while remaining in the same area, another cell pattern was observed (right panel) (and covered again with collagen). Figure Legend:
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