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. 2019 Jul 3;9:9622. doi: 10.1038/s41598-019-46075-1

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© The Author(s) 2019

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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Macrophages do not significantly alter spontaneous lipid vesicle formation in 3D-cultured orbital fibroblasts. (a) Representative Oil-Red-O (ORO) staining of lipid-containing vesicles in orbital fibroblasts in 3D culture for 7 days: HO2 without macrophages (left) and CO6 with macrophages (right, arrow), at a 1:1 fibroblast:macrophage ratio. Scale bar, 25 μm. (b) Macrophages did not alter spontaneous lipid vesicle formation in 3D culture. GO (HOs) or control (COs) fibroblasts (0.74 × 10⁵ cells/ml) were cultured with macrophages at a 1:1 fibroblast:macrophage ratio in the presence of 10% serum for 7 days. The fraction of ORO positive cells is shown as mean ± SEM (n = 3, no statistical difference between with and without macrophages). (c) ORO positive cells in control fibroblast lines, CO4 and CO7, did not significantly increase as the number of macrophages increased (mean ± SEM; n = 4, p = 0.118 (CO4) and p = 0.123 (CO7), Kruskal-Wallis test).