Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems

Background. In vitro methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) are a matter of priority for the in-depth research into the mechanisms of early embryogenesis. So-far, published results regarding the generation of hematopoietic cells come from studies using eithe...

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Autores principales:	German Atzin Mora-Roldan, Dalia Ramirez-Ramirez, Rosana Pelayo, Karlen Gazarian
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	hPSC primitive hematopoietic differentiation Wnt signaling embryoid body 3D culture Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/301aa97facf4473fa65b265e1e903858
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spelling	oai:doaj.org-article:301aa97facf4473fa65b265e1e9038582021-11-25T17:08:07ZAssessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems10.3390/cells101128582073-4409https://doaj.org/article/301aa97facf4473fa65b265e1e9038582021-10-01T00:00:00Zhttps://www.mdpi.com/2073-4409/10/11/2858https://doaj.org/toc/2073-4409Background. In vitro methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) are a matter of priority for the in-depth research into the mechanisms of early embryogenesis. So-far, published results regarding the generation of hematopoietic cells come from studies using either 2D or 3D culture formats, hence, it is difficult to discern their particular contribution to the development of the concept of a unique in vitro model in close resemblance to in vivo hematopoiesis. Aim of the study. To assess using the same culture conditions and the same time course, the potential of each of these two formats to support differentiation of human pluripotent stem cells to primitive hematopoiesis without exogenous activation of Wnt signaling. Methods. We used in parallel 2D and 3D formats, the same culture environment and assay methods (flow cytometry, IF, qPCR) to investigate stages of commitment and specification of mesodermal, and hemogenic endothelial cells to CD34 hematopoietic cells and evaluated their clonogenic capacity in a CFU system. Results. We show an adequate formation of mesoderm, an efficient commitment to hemogenic endothelium, a higher number of CD34 hematopoietic cells, and colony-forming capacity potential only in the 3D format-supported differentiation. Conclusions. This study shows that the 3D but not the 2D format ensures the induction and realization by endogenous mechanisms of human pluripotent stem cells’ intrinsic differentiation program to primitive hematopoietic cells. We propose that the 3D format provides an adequate level of upregulation of the endogenous Wnt/β-catenin signaling.German Atzin Mora-RoldanDalia Ramirez-RamirezRosana PelayoKarlen GazarianMDPI AGarticlehPSCprimitive hematopoietic differentiationWnt signalingembryoid body3D cultureBiology (General)QH301-705.5ENCells, Vol 10, Iss 2858, p 2858 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	hPSC primitive hematopoietic differentiation Wnt signaling embryoid body 3D culture Biology (General) QH301-705.5
spellingShingle	hPSC primitive hematopoietic differentiation Wnt signaling embryoid body 3D culture Biology (General) QH301-705.5 German Atzin Mora-Roldan Dalia Ramirez-Ramirez Rosana Pelayo Karlen Gazarian Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
description	Background. In vitro methods for hematopoietic differentiation of human pluripotent stem cells (hPSC) are a matter of priority for the in-depth research into the mechanisms of early embryogenesis. So-far, published results regarding the generation of hematopoietic cells come from studies using either 2D or 3D culture formats, hence, it is difficult to discern their particular contribution to the development of the concept of a unique in vitro model in close resemblance to in vivo hematopoiesis. Aim of the study. To assess using the same culture conditions and the same time course, the potential of each of these two formats to support differentiation of human pluripotent stem cells to primitive hematopoiesis without exogenous activation of Wnt signaling. Methods. We used in parallel 2D and 3D formats, the same culture environment and assay methods (flow cytometry, IF, qPCR) to investigate stages of commitment and specification of mesodermal, and hemogenic endothelial cells to CD34 hematopoietic cells and evaluated their clonogenic capacity in a CFU system. Results. We show an adequate formation of mesoderm, an efficient commitment to hemogenic endothelium, a higher number of CD34 hematopoietic cells, and colony-forming capacity potential only in the 3D format-supported differentiation. Conclusions. This study shows that the 3D but not the 2D format ensures the induction and realization by endogenous mechanisms of human pluripotent stem cells’ intrinsic differentiation program to primitive hematopoietic cells. We propose that the 3D format provides an adequate level of upregulation of the endogenous Wnt/β-catenin signaling.
format	article
author	German Atzin Mora-Roldan Dalia Ramirez-Ramirez Rosana Pelayo Karlen Gazarian
author_facet	German Atzin Mora-Roldan Dalia Ramirez-Ramirez Rosana Pelayo Karlen Gazarian
author_sort	German Atzin Mora-Roldan
title	Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
title_short	Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
title_full	Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
title_fullStr	Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
title_full_unstemmed	Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems
title_sort	assessment of the hematopoietic differentiation potential of human pluripotent stem cells in 2d and 3d culture systems
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/301aa97facf4473fa65b265e1e903858
work_keys_str_mv	AT germanatzinmoraroldan assessmentofthehematopoieticdifferentiationpotentialofhumanpluripotentstemcellsin2dand3dculturesystems AT daliaramirezramirez assessmentofthehematopoieticdifferentiationpotentialofhumanpluripotentstemcellsin2dand3dculturesystems AT rosanapelayo assessmentofthehematopoieticdifferentiationpotentialofhumanpluripotentstemcellsin2dand3dculturesystems AT karlengazarian assessmentofthehematopoieticdifferentiationpotentialofhumanpluripotentstemcellsin2dand3dculturesystems
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Assessment of the Hematopoietic Differentiation Potential of Human Pluripotent Stem Cells in 2D and 3D Culture Systems

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