Differentiation of monocytes and polarized M1/M2 macrophages from human induced pluripotent stem cells

Tea Soon Park; Rishabh Hirday; Russell Quinn; Sheela Panicker Jacob; Ricardo A. Feldman; Devika Bose; Ruchi Sharma; Kapil Bharti

STAR Protocols (Mar 2024)

Differentiation of monocytes and polarized M1/M2 macrophages from human induced pluripotent stem cells

Tea Soon Park,
Rishabh Hirday,
Russell Quinn,
Sheela Panicker Jacob,
Ricardo A. Feldman,
Devika Bose,
Ruchi Sharma,
Kapil Bharti

Affiliations

Tea Soon Park: Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA; Corresponding author
Rishabh Hirday: Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA
Russell Quinn: Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA
Sheela Panicker Jacob: Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA; Vita Therapeutics Inc., Baltimore, MD 21201, USA
Ricardo A. Feldman: Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA
Devika Bose: Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA
Ruchi Sharma: Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA
Kapil Bharti: Ocular Stem Cells and Translational Research (OSCTR) Section, Ophthalmic Genetic and Visual Function Branch (OGVFB), National Eye Institute, National Institutes of Health, Bethesda, MD 20892, USA; Corresponding author

Journal volume & issue: Vol. 5, no. 1
p. 102827

Abstract

Read online

Summary: Here, we present a protocol to differentiate induced pluripotent stem cell (iPSC) into adherent hematopoietic progenitors that release floating CD14+ CD45+ monocytes into the culture medium. We describe steps for iPSC expansion, embryoid body (EB) formation, suspension culture, plating EBs, and recurring harvests of monocytes, a.k.a. “monocyte factory.” We then describe detailed procedures for freezing/thawing of monocytes and differentiation into polarized M1 and M2 macrophages. This protocol provides foundation to study iPSC monocytes and their progenies such as macrophages, microglial, and dendritic cells.For complete details on the use and execution of this protocol, please refer to Karlson et al.1 and Panicker et al.2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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/

About the journal

Abstract

Keywords