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. 2019 Aug 22;9:12286. doi: 10.1038/s41598-019-48663-7

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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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The composition of immune infiltrates in tumour organoid derived isografts. (a) Pie charts showing percentages of granulocytes (CD45⁺CD11b⁺Ly6G⁺), macrophages (CD45⁺CD11b⁺F4/80⁺), B cells (CD45⁺B220⁺), cytotoxic T cells (CD45⁺CD3⁺CD8⁺), and T helper lymphocytes (CD45⁺CD3⁺CD4⁺) in the pancreatic tissues from mice at 2 months (left), 4 months (middle) and 6 months (right) from organoid transplantation. “Unstained” denotes pancreatic epithelial cells and other stromal components. Colour key is provided. (b) Pie charts showing the immune cell populations defined in (a) as percentages of CD45⁺ cells. Colour key as in (a). (c) Flow cytometry analysis of the indicated CD45⁺ cell populations (granulocytes and macrophages) in the pancreas of preinvasive, classical tumours, or PDC from (a). (d) Flow cytometry analysis of the indicated immune cell populations (granulocytes and macrophages) in the pancreas of preinvasive, classical tumours, or PDC from (b). (e) Flow cytometry analysis of the indicated CD45⁺ cell populations (T cells) in the pancreas of preinvasive, classical tumours, or PDC from (a). (f) Flow cytometry analysis of the indicated immune cell populations (T cells) in the pancreas of preinvasive, classical tumours, or PDC from (b). Each dot in the graphics (c–f) refers to the individual tumour samples available for cytometric evaluation. ODIs from B6-K1, B6-K2, and B6-K3 organoids are identified by circles, triangles, and squares, respectively. (g) Immunohistochemical staining for CD8 in tissue from mice bearing preinvasive lesions (B6-K2, left), classical tumours (B6-K2, middle) or poorly differentiated tumours (B6-K3, right). Scale bars, 50 µm. Quantification is provided on the left as the average number of CD8 positive cells per mm² in preinvasive (n = 6), classical (n = 10), and poorly differentiated (PDC, n = 8) tumours. From 3 to 5 individual areas per case were examined. Statistical associations were determined by Student’s t-test. *p < 0.05; **p < 0.01; ***p < 0.001. See also Supplementary Fig. 2 for details.