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

doi:10.1016/j.xpro.2023.102827

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

STAR Protoc. 2024 Mar 15;5(1):102827. doi: 10.1016/j.xpro.2023.102827. Epub 2024 Jan 13.

Authors

Tea Soon Park¹, Rishabh Hirday², Russell Quinn², Sheela Panicker Jacob³, Ricardo A Feldman⁴, Devika Bose², Ruchi Sharma², Kapil Bharti⁵

Affiliations

¹ 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. Electronic address: teasoon.park@nih.gov.
² 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.
³ Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA; Vita Therapeutics Inc., Baltimore, MD 21201, USA.
⁴ Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201, USA.
⁵ 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. Electronic address: kapil.bharti@nih.gov.

Abstract

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.

Keywords: Cell Biology; Cell Differentiation; Stem Cells.

MeSH terms

Cell Differentiation
Embryoid Bodies
Humans
Induced Pluripotent Stem Cells*
Macrophages
Monocytes*