Protocol for differentiation of monocytes and macrophages from human induced pluripotent stem cells

Kevin Emmerich; Francesca Calcaterra; Xuming Tang; Guibin Chen; Elena Pontarini; Roberta Ciceri; Dan Yang; Benjamin Joseph; Silvia Della Bella; Isabella Varea; Domenico Mavilio; Manfred Boehm

doi:10.1016/j.xpro.2024.103217

Protocol for differentiation of monocytes and macrophages from human induced pluripotent stem cells

STAR Protoc. 2024 Sep 20;5(3):103217. doi: 10.1016/j.xpro.2024.103217. Epub 2024 Jul 27.

Affiliations

¹ Translational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20814, USA. Electronic address: emmerichkb@nih.gov.
² Unit of Clinical and Experimental Immunology, IRCCS Humanitas Research Hospital, 20089 Rozzano, Italy; Department of Medical Biotechnologies and Translational Medicine (BioMeTra), University of Milan, 20054 Segrate, Italy.
³ Translational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20814, USA.
⁴ Unit of Clinical and Experimental Immunology, IRCCS Humanitas Research Hospital, 20089 Rozzano, Italy; Center for Experimental Medicine and Rheumatology, William Harvey Research Institute, London, UK.
⁵ Translational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20814, USA. Electronic address: boehmm@nih.gov.

Abstract

Study of disease-relevant immune cells, namely monocytes and macrophages, is limited based on availability of primary tissue, a limitation that can be remedied using human induced pluripotent stem cell (hiPSC) technology. Here, we present a protocol for differentiation of monocytes and macrophages from hiPSCs. We describe steps for hiPSC maintenance, mesoderm lineage induction, hematopoietic progenitor cells (HPCs) commitment and expansion, and myeloid lineage induction. We then detail procedures for monocyte formation and functional macrophage formation and polarization. For complete details on the use and execution of this protocol, please refer to Chen et al.¹.

Keywords: Cell Biology; Cell Differentiation; Flow Cytometry; Immunology; Microscopy; Stem Cells.

MeSH terms

Cell Culture Techniques / methods
Cell Differentiation* / physiology
Cells, Cultured
Hematopoietic Stem Cells / cytology
Humans
Induced Pluripotent Stem Cells* / cytology
Macrophages* / cytology
Monocytes* / cytology

Grants and funding

ZIA HL006079/ImNIH/Intramural NIH HHS/United States