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Stem Cell Res. 2019 Oct 15;41:101600. doi: 10.1016/j.scr.2019.101600. [Epub ahead of print]

Robust generation of erythroid and multilineage hematopoietic progenitors from human iPSCs using a scalable monolayer culture system.

Author information

1
Cellular and Molecular Therapeutics Branch, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health (NIH), 9000 Rockville, Bethesda, MD 20892, United States.
2
Translational Vascular Medicine Branch, NHLBI, NIH, Bethesda, MD 20892, United States.
3
Clinical Flow Core Facility, NHLBI, NIH, Bethesda, MD 20892, United States.
4
Transgenic Core Facility, NHLBI, NIH, Bethesda, MD 20892, United States.
5
iPSC Core Facility, NHLBI, NIH, Bethesda, MD 20892, United States.
6
Cellular and Molecular Therapeutics Branch, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health (NIH), 9000 Rockville, Bethesda, MD 20892, United States; College of Applied Medical Sciences, University of Hail, Hail, Saudi Arabia; Department of Biology, The Catholic University of America, Washington, DC 20064, United States.
7
Cellular and Molecular Therapeutics Branch, National Heart, Lung and Blood Institute (NHLBI), National Institutes of Health (NIH), 9000 Rockville, Bethesda, MD 20892, United States. Electronic address: larochea@nhlbi.nih.gov.

Abstract

One of the most promising objectives of clinical hematology is to derive engraftable autologous hematopoietic stem cells (HSCs) from human induced pluripotent stem cells (iPSCs). Progress in translating iPSC technologies to the clinic relies on the availability of scalable differentiation methodologies. In this study, human iPSCs were differentiated for 21 days using STEMdiff™, a monolayer-based approach for hematopoietic differentiation of human iPSCs that requires no replating, co-culture or embryoid body formation. Both hematopoietic and non-hematopoietic cells were functionally characterized throughout differentiation. In the hematopoietic fraction, an early transient population of primitive CD235a+ erythroid progenitor cells first emerged, followed by hematopoietic progenitors with multilineage differentiation activity in vitro but no long-term engraftment potential in vivo. In later stages of differentiation, a nearly exclusive production of definitive erythroid progenitors was observed. In the non-hematopoietic fraction, we identified a prevalent population of mesenchymal stromal cells and limited arterial vascular endothelium (VE), suggesting that the cellular constitution of the monolayer may be inadequate to support the generation of HSCs with durable repopulating potential. Quantitative modulation of WNT/β-catenin and activin/nodal/TGFβ signaling pathways with CHIR/SB molecules during differentiation enhanced formation of arterial VE, definitive multilineage and erythroid progenitors, but was insufficient to orchestrate the generation of engrafting HSCs. Overall, STEMdiff™ provides a clinically-relevant and readily adaptable platform for the generation of erythroid and multilineage hematopoietic progenitors from human pluripotent stem cells.

KEYWORDS:

Arterial vascular endothelium; Erythroid progenitor; Hematopoietic stem and progenitor cell; Induced pluripotent stem cell; Monolayer differentiation system; STEMdiffTM hematopoietic differentiation kit

PMID:
31710911
DOI:
10.1016/j.scr.2019.101600
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