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Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor

BACKGROUND: hPSC-derived endothelial and hematopoietic cells (ECs and HCs) are an interesting source of cells for tissue engineering. Despite their close spatial and temporal embryonic development, current hPSC differentiation protocols are specialized in only one of these lineages. In this study, w...

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Autores principales: Vargas-Valderrama, Alejandra, Ponsen, Anne-Charlotte, Le Gall, Morgane, Clay, Denis, Jacques, Sébastien, Manoliu, Tudor, Rouffiac, Valérie, Ser-le-Roux, Karine, Quivoron, Cyril, Louache, Fawzia, Uzan, Georges, Mitjavila-Garcia, Maria-Teresa, Oberlin, Estelle, Guenou, Hind
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9205076/
https://www.ncbi.nlm.nih.gov/pubmed/35715824
http://dx.doi.org/10.1186/s13287-022-02925-w
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author Vargas-Valderrama, Alejandra
Ponsen, Anne-Charlotte
Le Gall, Morgane
Clay, Denis
Jacques, Sébastien
Manoliu, Tudor
Rouffiac, Valérie
Ser-le-Roux, Karine
Quivoron, Cyril
Louache, Fawzia
Uzan, Georges
Mitjavila-Garcia, Maria-Teresa
Oberlin, Estelle
Guenou, Hind
author_facet Vargas-Valderrama, Alejandra
Ponsen, Anne-Charlotte
Le Gall, Morgane
Clay, Denis
Jacques, Sébastien
Manoliu, Tudor
Rouffiac, Valérie
Ser-le-Roux, Karine
Quivoron, Cyril
Louache, Fawzia
Uzan, Georges
Mitjavila-Garcia, Maria-Teresa
Oberlin, Estelle
Guenou, Hind
author_sort Vargas-Valderrama, Alejandra
collection PubMed
description BACKGROUND: hPSC-derived endothelial and hematopoietic cells (ECs and HCs) are an interesting source of cells for tissue engineering. Despite their close spatial and temporal embryonic development, current hPSC differentiation protocols are specialized in only one of these lineages. In this study, we generated a hematoendothelial population that could be further differentiated in vitro to both lineages. METHODS: Two hESCs and one hiPSC lines were differentiated into a hematoendothelial population, hPSC-ECs and blast colonies (hPSC-BCs) via CD144(+)-embryoid bodies (hPSC-EBs). hPSC-ECs were characterized by endothelial colony-forming assay, LDL uptake assay, endothelial activation by TNF-α, nitric oxide detection and Matrigel-based tube formation. Hematopoietic colony-forming cell assay was performed from hPSC-BCs. Interestingly, we identified a hPSC-BC population characterized by the expression of both CD144 and CD45. hPSC-ECs and hPSC-BCs were analyzed by flow cytometry and RT-qPCR; in vivo experiments have been realized by ischemic tissue injury model on a mouse dorsal skinfold chamber and hematopoietic reconstitution in irradiated immunosuppressed mouse from hPSC-ECs and hPSC-EB-CD144(+), respectively. Transcriptomic analyses were performed to confirm the endothelial and hematopoietic identity of hESC-derived cell populations by comparing them against undifferentiated hESC, among each other’s (e.g. hPSC-ECs vs. hPSC-EB-CD144(+)) and against human embryonic liver (EL) endothelial, hematoendothelial and hematopoietic cell subpopulations. RESULTS: A hematoendothelial population was obtained after 84 h of hPSC-EBs formation under serum-free conditions and isolated based on CD144 expression. Intrafemorally injection of hPSC-EB-CD144(+) contributed to the generation of CD45(+) human cells in immunodeficient mice suggesting the existence of hemogenic ECs within hPSC-EB-CD144(+). Endothelial differentiation of hPSC-EB-CD144(+) yields a population of > 95% functional ECs in vitro. hPSC-ECs derived through this protocol participated at the formation of new vessels in vivo in a mouse ischemia model. In vitro, hematopoietic differentiation of hPSC-EB-CD144(+) generated an intermediate population of > 90% CD43(+) hPSC-BCs capable to generate myeloid and erythroid colonies. Finally, the transcriptomic analyses confirmed the hematoendothelial, endothelial and hematopoietic identity of hPSC-EB-CD144(+), hPSC-ECs and hPSC-BCs, respectively, and the similarities between hPSC-BC-CD144(+)CD45(+), a subpopulation of hPSC-BCs, and human EL hematopoietic stem cells/hematopoietic progenitors. CONCLUSION: The present work reports a hPSC differentiation protocol into functional hematopoietic and endothelial cells through a hematoendothelial population. Both lineages were proven to display characteristics of physiological human cells, and therefore, they represent an interesting rapid source of cells for future cell therapy and tissue engineering. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-022-02925-w.
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spelling pubmed-92050762022-06-18 Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor Vargas-Valderrama, Alejandra Ponsen, Anne-Charlotte Le Gall, Morgane Clay, Denis Jacques, Sébastien Manoliu, Tudor Rouffiac, Valérie Ser-le-Roux, Karine Quivoron, Cyril Louache, Fawzia Uzan, Georges Mitjavila-Garcia, Maria-Teresa Oberlin, Estelle Guenou, Hind Stem Cell Res Ther Research BACKGROUND: hPSC-derived endothelial and hematopoietic cells (ECs and HCs) are an interesting source of cells for tissue engineering. Despite their close spatial and temporal embryonic development, current hPSC differentiation protocols are specialized in only one of these lineages. In this study, we generated a hematoendothelial population that could be further differentiated in vitro to both lineages. METHODS: Two hESCs and one hiPSC lines were differentiated into a hematoendothelial population, hPSC-ECs and blast colonies (hPSC-BCs) via CD144(+)-embryoid bodies (hPSC-EBs). hPSC-ECs were characterized by endothelial colony-forming assay, LDL uptake assay, endothelial activation by TNF-α, nitric oxide detection and Matrigel-based tube formation. Hematopoietic colony-forming cell assay was performed from hPSC-BCs. Interestingly, we identified a hPSC-BC population characterized by the expression of both CD144 and CD45. hPSC-ECs and hPSC-BCs were analyzed by flow cytometry and RT-qPCR; in vivo experiments have been realized by ischemic tissue injury model on a mouse dorsal skinfold chamber and hematopoietic reconstitution in irradiated immunosuppressed mouse from hPSC-ECs and hPSC-EB-CD144(+), respectively. Transcriptomic analyses were performed to confirm the endothelial and hematopoietic identity of hESC-derived cell populations by comparing them against undifferentiated hESC, among each other’s (e.g. hPSC-ECs vs. hPSC-EB-CD144(+)) and against human embryonic liver (EL) endothelial, hematoendothelial and hematopoietic cell subpopulations. RESULTS: A hematoendothelial population was obtained after 84 h of hPSC-EBs formation under serum-free conditions and isolated based on CD144 expression. Intrafemorally injection of hPSC-EB-CD144(+) contributed to the generation of CD45(+) human cells in immunodeficient mice suggesting the existence of hemogenic ECs within hPSC-EB-CD144(+). Endothelial differentiation of hPSC-EB-CD144(+) yields a population of > 95% functional ECs in vitro. hPSC-ECs derived through this protocol participated at the formation of new vessels in vivo in a mouse ischemia model. In vitro, hematopoietic differentiation of hPSC-EB-CD144(+) generated an intermediate population of > 90% CD43(+) hPSC-BCs capable to generate myeloid and erythroid colonies. Finally, the transcriptomic analyses confirmed the hematoendothelial, endothelial and hematopoietic identity of hPSC-EB-CD144(+), hPSC-ECs and hPSC-BCs, respectively, and the similarities between hPSC-BC-CD144(+)CD45(+), a subpopulation of hPSC-BCs, and human EL hematopoietic stem cells/hematopoietic progenitors. CONCLUSION: The present work reports a hPSC differentiation protocol into functional hematopoietic and endothelial cells through a hematoendothelial population. Both lineages were proven to display characteristics of physiological human cells, and therefore, they represent an interesting rapid source of cells for future cell therapy and tissue engineering. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13287-022-02925-w. BioMed Central 2022-06-17 /pmc/articles/PMC9205076/ /pubmed/35715824 http://dx.doi.org/10.1186/s13287-022-02925-w Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://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.
spellingShingle Research
Vargas-Valderrama, Alejandra
Ponsen, Anne-Charlotte
Le Gall, Morgane
Clay, Denis
Jacques, Sébastien
Manoliu, Tudor
Rouffiac, Valérie
Ser-le-Roux, Karine
Quivoron, Cyril
Louache, Fawzia
Uzan, Georges
Mitjavila-Garcia, Maria-Teresa
Oberlin, Estelle
Guenou, Hind
Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor
title Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor
title_full Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor
title_fullStr Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor
title_full_unstemmed Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor
title_short Endothelial and hematopoietic hPSCs differentiation via a hematoendothelial progenitor
title_sort endothelial and hematopoietic hpscs differentiation via a hematoendothelial progenitor
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9205076/
https://www.ncbi.nlm.nih.gov/pubmed/35715824
http://dx.doi.org/10.1186/s13287-022-02925-w
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