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. 2020 Jan 16;10:474. doi: 10.1038/s41598-019-57173-5

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© The Author(s) 2020

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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Heparin administration in stored blood leads to extracellular DNA deposition on the outlet of the filter. (A) Citrated blood (Cit-blood) stored for 0, 5, or 7 days was administered to the circuit and then 5 U/mL of unfractionated heparin was administered at 15 minutes. In-circuit pressure was monitored for 60 minutes. N = 4 per group. The paired t-test was used to analyze the difference between in-circuit pressure at 15 and 60 minutes. *p < 0.05, **p < 0.01. (B) The filter was removed at 60 minutes and analyzed by immunofluorescence. Fibrin(ogen) was labeled in red and DNA was labeled in blue. Whole images of the filters (scale bars: 10 mm) were analyzed by BZ-X700 microscopy and magnified images (scale bars: 50 μm) were analyzed by LSM 700 confocal microscopy. Representative images of N = 5 are shown.