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. 2024 Jul 6;15:199. doi: 10.1186/s13287-024-03817-x

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 7 — Potential of h-NG2/_BMMSCs to develop BDCs and their unique ability to directly differentiate into sinusoidal cells and structures in response to DEN-induced liver injury cues. (Aa-c) During 1–4 h of culture in _DENCM, BDC-like morphological changes were observed in culturing h-NG2/_BMMSC cells that differentiated from h-NG2/_BMMSCs (a, red arrows, box), but few similar changes were observed in cells that differentiated from parental h-_BMMSCs (b, box), and the number of changed cells per image field was analyzed (c, n = 10). (Ba, b) Double IF staining of CK19⁺ cells (a, red, second panels) covered with h-NG2/_BMMSCs (NG2, green, top panel) or h-_BMMSCs (CD90, green, top panel) and analysis of the percentage of developed CK19⁺ cells/image half-field from merged cells (b, brown, boxes) during 18–24 h of culture in _DENCM, n = 6. (Ca-c) Double IF staining revealed that during the 18–24 h period in _DENCM cultures, the CK19⁺ cells developmed from h-NG2/_BMMSCs formed obvious vessel-like structures (a, merged, arrows) that were not observed in the parental h-_BMMSCs (b), and quantification was performed (c, n = 6). (Da-c) Double IF staining of h-NG2/_BMMSCs (NG2, green, a) and h-_BMMSCs (CD90, green, b) covered with Lyve-1⁺ cells (red) after 4–6 h culture periods in _DENCM and quantification of the merged cells in Da, b (c, boxes), n = 6. (Ea, b) In the _DENCM cultures, Lyve-1⁺ cell (red)-generated from h-NG2/_BMMSC cells (green) formed vessel-like structures (a, arrows) and this phenomena was not appeared in the h-_BMMSCs (b). (F) Quantification of the data in E (n = 6). Scale bar = 200 μm for all images. At least three independent experiments were performed, and the data are presented as the means ± SDs; **p < 0.001 compared with h-_BMMSCs