Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA

Aditi Majumder; Kran Suknuntha; David Bennin; Lucas Klemm; Vera S. Brok-Volchanskaya; Anna Huttenlocher; Igor Slukvin

STAR Protocols (Sep 2020)

Generation of Human Neutrophils from Induced Pluripotent Stem Cells in Chemically Defined Conditions Using ETV2 Modified mRNA

Aditi Majumder,
Kran Suknuntha,
David Bennin,
Lucas Klemm,
Vera S. Brok-Volchanskaya,
Anna Huttenlocher,
Igor Slukvin

Affiliations

Aditi Majumder: Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA
Kran Suknuntha: Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA; Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI 53792, USA; Chakri Naruebodindra Medical Institute, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Samut Prakan 10540, Thailand
David Bennin: Chakri Naruebodindra Medical Institute, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Samut Prakan 10540, Thailand
Lucas Klemm: Department of Pediatrics and Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA
Vera S. Brok-Volchanskaya: Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA
Anna Huttenlocher: Department of Pediatrics and Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, WI 53706, USA
Igor Slukvin: Wisconsin National Primate Research Center, University of Wisconsin, Madison, WI 53715, USA; Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison, WI 53792, USA; Department of Cell and Regenerative Biology, University of Wisconsin School of Medicine and Public Health, Madison, WI 53707-7365, USA; Corresponding author

Journal volume & issue: Vol. 1, no. 2
p. 100075

Abstract

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Summary: This protocol describes a rapid and efficient feeder-, serum-, and xeno-free method for neutrophil generation from hiPSCs using ETV2 modified mRNA (mmRNA), which directs hematoendothelial programming of hiPSCs. Hematoendothelial progenitors were cultured with GM-CSF, FGF-2, and UM171 to expand myelomonocytic progenitors, followed by treatment with G-CSF and retinoic acid agonist Am580 to induce neutrophil maturation. This protocol is suitable for generating functional neutrophils from iPSCs to interrogate the role of genes in a neutrophil development and function.For complete details on the use and execution of this protocol, please refer to Brok-Volchanskaya et al. (2019).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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