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. 2020 Jun 3;13:63. doi: 10.1186/s13045-020-00899-x

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© The Author(s) 2020

Open AccessThis 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. 1 — Isolation and expansion of CD90-CD13-CD44+MCSCs from an AML patient bone marrow ex vivo. a Phase bright images of MSCs and MCSCs at different passages (P). b 24 h after culturing cells with BrdU, P7 MSCs, and P7 MCSCs were collected and analyzed by FACS for incorporation of BrdU. c Aggregate data showing percentage of BrdU+CD44+ cells in MSCs and MCSCs. d P3 MSCs and P3 MCSCs were analyzed by FACS for immunophenotype. Thick black arrows indicate gating strategy. The red arrow indicates the population of CD90-CD13-CD44+ cells. e Aggregate FACS data showing the percentage of MSCs and MCSCs positive for each biomarker. f Left: Phase bright image of floating cells from P5 MSC culture. Right: P5 MSCs were collected and analyzed by FACS. g Left: Phase bright image of floating cells from P5 MCSC culture. Right: P5 MCSCs were collected and analyzed by FACS. The red arrow indicates CD90-CD13-CD44+ cells. hAggregate FACS data showing the percentage of CD90-CD13-CD44+ cells comprising the floating cell populations from MSCs and MCSCs. iCumulative counts of clusters generated from MSC and MCSC cultures over time. j Inset: phase bright images of floating tube-like structure from P8 MCSC culture. Black thick arrows indicate the same round tube at × 2 and × 10 magnification. k P8 Floating MCSCs were analyzed by FACS for endothelial cell markers such as VE-Cadherin (CD144). l Phase bright image of GFP+MOLM-14 cells cultured with MSCs after 24-h plating. m Phase bright image of GFP+MOLM-14 cells co-cultured with MCSCs after 24-h plating. Where applicable, data are means ± SEM from each group and were analyzed by Student t test. *p < 0.05; N=3. Scale bar 100 μm