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. 2018 Mar 1;8:3852. doi: 10.1038/s41598-018-21610-8

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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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hMAPCs support lymph node survival and reconnection through blood vessel and lymphatic collector regeneration. (a–c) Bright field images of blood vascular network leading up to the transplanted lymph node (LN) of mice treated with Matrigel containing PBS (a) or hMAPCs (‘hM’; b) 16 weeks (w) earlier, and corresponding quantification (c; n = 6; *P = 0.0011 versus PBS by unpaired two-tailed Student’s t-test). (d) Merged bright field/fluorescence image of an eGFP⁺ (in green) LN 16 w post-transplantion and treatment with Matrigel containing hMAPCs revealing that the LN is irrigated by numerous blood vessels. (e,f) Merged bright field and fluorescence images zooming in on a DsRed⁺ (in red) LN 8 w post-transplantion and treatment with Matrigel containing hMAPCs revealing extensive LN vascular network branching. Inset (i1) corresponds to boxed area in f. (g) Merged immunofluorescence image of a Prox1 (in red)/eGFP (in green)-stained section in a mouse treated with Matrigel + hMAPCs 16 w earlier revealing that part of the branches are lymphatic in nature (Prox1⁺, indicated by arrowheads). Inset (i2) corresponds to boxed area in g. (h–j) LYVE1-stained (in red) cross-sections of PBS (h) or hMAPC-treated (‘hM’; i) mice in the area around the sutures at 8 w after LN transplantation and corresponding quantification (j; n = 5–8; *P = 0.0014 by unpaired two-tailed Student’s t-test versus PBS). (k–n) Fluorescence images of the area around the transplanted eGFP⁺ LN (in green; lined by a dashed line in k; adjacent Prox1 (in green)-stained section is shown in l; Prox1 (in green)/αSMA (in red) (arrowheads; double staining in m zooms in on the boxed area in k,l; and n represents the same area on an adjacent LYVE1 (in red)-stained cross-section) revealing Prox1⁺αSMA⁺LYVE1^- lymphatic collector vessels in mice treated 16 w earlier with Matrigel containing hMAPCs. Asterisks in l-n indicate lymph (which artefactually fluoresces upon exposure to tyramide-based amplification which was used for Prox1 and LYVE1 staining). White arrows in a, b, e-i, k, l indicate sutures used to fix the transplanted LN.