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. 2023 Sep 12;12(1):2240670. doi: 10.1080/2162402X.2023.2240670

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© 2023 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent.

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Figure 2. — Hematopoietic Progenitor Cell Characterization and Transplantation Experiments.

a. Phase contrast (PhC) image of monolayer progenitor population at Day 5 of differentiation. b. PhC image showing a vessel within the vascular organoid. c. Immunohistochemistry image showing the vascular organoid. Hematopoietic progenitor cells expressing CD43 develop inside of a tubular structure. d. The tubular structures of the organoid are formed by endothelial cells (stained for VE-Cadherin). e. Heatmap showing significant up-regulation in endothelial and hematopoietic gene expression of Day 5 progenitors, compared to hPSC control. f. ShinyGO functional enrichment tool showing that the most significantly up-regulated genes in Day 5 progenitors are predominantly affiliated with hematopoietic processes. g. Flow cytometry images demonstrating that cells from Day 5 of CHIR differentiation exhibit a CD34⁺CD90⁺CD73⁻ engraftment signature. h. Flow cytometric analysis, conducted at 8 weeks post-injection, showing the percentage of human CD45⁺ cells generated by hematopoietic progenitor cells at time point 1 (HPC1). i. Flow cytometric analysis, conducted at 21 weeks post-injection, showing the percentage of human CD45⁺ cells generated by HPC1. j. Flow cytometric analysis, conducted at 8 weeks post-injection, showing the percentage of human CD45⁺ cells generated by hematopoietic progenitor cells at time point 2 (HPC2). k. Flow cytometric analysis, conducted at 12 weeks post-injection, showing the percentage of human CD45⁺ cells generated by hematopoietic progenitor cells at time point 3 (HPC3). l. A graph demonstrating a trend toward an increasing percentage of engrafted HPC2 (from week 8 to week 13). Bars represent mean ± SEM from at least three independent experiments. m. A graph demonstrating that there is no significant increase in engraftment potential at time point HPC3 compared to HPC2. Bars represent mean ± SEM from at least three independent experiments. o. A graph demonstrating no significant increase in engraftment potential when increasing the number of injected cells from 0.5 million to 2 million. Bars represent mean ± SEM from at least three independent experiments.