Differentiation of Human Induced Pluripotent Stem Cell into Macrophages

As a model to interrogate human macrophage biology, macrophages differentiated from human induced pluripotent stem cells (hiPSCs) transcend other existing models by circumventing the variability seen in human monocyte-derived macrophages, whilst epitomizing macrophage phenotypic and functional chara...

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Veröffentlicht in:Bio-protocol 2022-03, Vol.12 (6), p.e4361-e4361
Hauptverfasser: Douthwaite, Harriet, Arteagabeitia, Aitor Bermejo, Mukhopadhyay, Subhankar
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Sprache:eng
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Zusammenfassung:As a model to interrogate human macrophage biology, macrophages differentiated from human induced pluripotent stem cells (hiPSCs) transcend other existing models by circumventing the variability seen in human monocyte-derived macrophages, whilst epitomizing macrophage phenotypic and functional characteristics over those offered by macrophage-like cell lines ( Mukherjee , 2018 ). Furthermore, hiPSCs are amenable to genetic manipulation, unlike human monocyte-derived macrophages (MDMs) (van Wilgenburg , 2013 ; Lopez- Yrigoyen , 2020 ), proposing boundless opportunities for specific disease modelling. We outline an effective and efficient protocol that delivers a continual production of hiPSC-derived-macrophages (iMACs), exhibiting human macrophage surface and intracellular markers, together with functional activity. The protocol describes the resuscitation, culture, and differentiation of hiPSC into mature terminal macrophages, via the initial and intermediate steps of expansion of hiPSCs, formation into embryoid bodies (EBs), and generation of hematopoietic myeloid precursors. We offer a simplified, scalable, and adaptable technique that advances upon other protocols, utilizing feeder-free conditions and reduced growth factors, to produce high yields of consistent iMACs over a period of several months, economically.
ISSN:2331-8325
2331-8325
DOI:10.21769/BioProtoc.4361