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Updated: Aug 2, 2025
Production and Characterization of Human Macrophages from Pluripotent Stem Cells
Published on: April 16, 2020
Application of human iPSC-derived macrophages in a miniaturized high-content-imaging-based efferocytosis assay
Sarah Bitzer1, Mozhgan Dehghan Harati1, Karim C El Kasmi2
1Department of Drug Discovery Sciences, Boehringer Ingelheim Pharma GmbH & Co. KG, 88397 Biberach an der Riss, Germany.
Abstract:
Macrophages play a pivotal role in drug discovery due to their key regulatory functions in health and disease. Overcoming the limited availability and donor variability of human monocyte-derived macrophages (MDMs), human induced pluripotent stem cell (iPSC)-derived macrophages (IDMs) could provide a promising tool for both disease modeling and drug discovery. To access large numbers of model cells for medium- to high-throughput application purposes, an upscaled protocol was established for differentiation of iPSCs into progenitor cells and subsequent maturation into functional macrophages. These IDM cells resembled MDMs both with respect to surface marker expression and phago- as well as efferocytotic function. A statistically robust high-content-imaging assay was developed to quantify the efferocytosis rate of IDMs and MDMs allowing for measurements both in the 384- and 1536-well microplate format. Validating the applicability of the assay, inhibitors of spleen tyrosine kinase (Syk) were shown to modulate efferocytosis in IDMs and MDMs with comparable pharmacology. The miniaturized cellular assay with the upscaled provision of macrophages opens new routes to pharmaceutical drug discovery in the context of efferocytosis-modulating substances.
Insights
Human induced pluripotent stem cell-derived macrophages (IDMs) offer a scalable alternative to donor-derived cells for drug discovery. A new assay effectively measures efferocytosis in IDMs, supporting the development of new therapeutics.
Area of Science:
- Cell Biology
- Immunology
- Drug Discovery
Background:
- Macrophages are crucial in health and disease, making them vital for drug discovery.
- Human monocyte-derived macrophages (MDMs) have limitations in availability and donor variability.
- Human induced pluripotent stem cell-derived macrophages (IDMs) present a potential solution for consistent cell sourcing.
Purpose of the Study:
- To develop an upscaled protocol for generating functional IDMs.
- To establish a robust high-content imaging assay for quantifying macrophage efferocytosis.
- To validate the assay's utility in drug discovery by testing spleen tyrosine kinase (Syk) inhibitors.
Main Methods:
- Upscaled differentiation of induced pluripotent stem cells (iPSCs) into macrophages.
- Characterization of IDM surface markers and phagocytic/efferocytotic functions.
- Development and validation of a high-content imaging assay for efferocytosis in 384- and 1536-well formats.
Main Results:
- IDMs exhibited functional and phenotypic similarities to MDMs.
- The developed assay accurately quantified efferocytosis rates in both IDMs and MDMs.
- Spleen tyrosine kinase (Syk) inhibitors demonstrated comparable pharmacological effects on efferocytosis in both cell types.
Conclusions:
- Upscaled IDM generation provides a consistent and abundant cell source for research.
- The miniaturized efferocytosis assay is suitable for medium- to high-throughput drug screening.
- This approach facilitates the discovery of novel efferocytosis-modulating drugs.
