Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA
© 2019 The Authors Human induced pluripotent stem cells (hiPSCs) can serve as a versatile and scalable source of neutrophils for biomedical research and transfusion therapies. Here we describe a rapid efficient serum- and xenogen-free protocol for neutrophil generation, which is based on direct hema...
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th-mahidol.499972020-01-27T14:34:34Z Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA Vera S. Brok-Volchanskaya David A. Bennin Kran Suknuntha Lucas C. Klemm Anna Huttenlocher Igor Slukvin University of Wisconsin-Madison University of Wisconsin School of Medicine and Public Health Mahidol University Wisconsin National Primate Research Center Biochemistry, Genetics and Molecular Biology © 2019 The Authors Human induced pluripotent stem cells (hiPSCs) can serve as a versatile and scalable source of neutrophils for biomedical research and transfusion therapies. Here we describe a rapid efficient serum- and xenogen-free protocol for neutrophil generation, which is based on direct hematoendothelial programming of hiPSCs using ETV2-modified mRNA. Culture of ETV2-induced hematoendothelial progenitors in the presence of GM-CSF, FGF2, and UM171 led to continuous production of generous amounts of CD34+CD33+ myeloid progenitors which could be harvested every 8–10 days for up to 30 days of culture. Subsequently, myeloid progenitors were differentiated into neutrophils in the presence of G-CSF and the retinoic acid agonist Am580. Neutrophils obtained in these conditions displayed a typical somatic neutrophil morphology, produced reactive oxygen species, formed neutrophil extracellular traps and possessed phagocytic and chemotactic activities. Overall, this technology offers an opportunity to generate a significant number of neutrophils as soon as 14 days after initiation of differentiation. 2020-01-27T07:34:34Z 2020-01-27T07:34:34Z 2019-12-10 Article Stem Cell Reports. Vol.13, No.6 (2019), 1099-1110 10.1016/j.stemcr.2019.10.007 22136711 2-s2.0-85075885519 https://repository.li.mahidol.ac.th/handle/123456789/49997 Mahidol University SCOPUS https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85075885519&origin=inward |
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Biochemistry, Genetics and Molecular Biology Vera S. Brok-Volchanskaya David A. Bennin Kran Suknuntha Lucas C. Klemm Anna Huttenlocher Igor Slukvin Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA |
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© 2019 The Authors Human induced pluripotent stem cells (hiPSCs) can serve as a versatile and scalable source of neutrophils for biomedical research and transfusion therapies. Here we describe a rapid efficient serum- and xenogen-free protocol for neutrophil generation, which is based on direct hematoendothelial programming of hiPSCs using ETV2-modified mRNA. Culture of ETV2-induced hematoendothelial progenitors in the presence of GM-CSF, FGF2, and UM171 led to continuous production of generous amounts of CD34+CD33+ myeloid progenitors which could be harvested every 8–10 days for up to 30 days of culture. Subsequently, myeloid progenitors were differentiated into neutrophils in the presence of G-CSF and the retinoic acid agonist Am580. Neutrophils obtained in these conditions displayed a typical somatic neutrophil morphology, produced reactive oxygen species, formed neutrophil extracellular traps and possessed phagocytic and chemotactic activities. Overall, this technology offers an opportunity to generate a significant number of neutrophils as soon as 14 days after initiation of differentiation. |
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University of Wisconsin-Madison |
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University of Wisconsin-Madison Vera S. Brok-Volchanskaya David A. Bennin Kran Suknuntha Lucas C. Klemm Anna Huttenlocher Igor Slukvin |
format |
Article |
author |
Vera S. Brok-Volchanskaya David A. Bennin Kran Suknuntha Lucas C. Klemm Anna Huttenlocher Igor Slukvin |
author_sort |
Vera S. Brok-Volchanskaya |
title |
Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA |
title_short |
Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA |
title_full |
Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA |
title_fullStr |
Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA |
title_full_unstemmed |
Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA |
title_sort |
effective and rapid generation of functional neutrophils from induced pluripotent stem cells using etv2-modified mrna |
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2020 |
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https://repository.li.mahidol.ac.th/handle/123456789/49997 |
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1763493723236204544 |