Efficient, long term production of monocyte-derived macrophages from human pluripotent stem cells under partly-defined and fully-defined conditions
Open accessAbstract
Human macrophages are specialised hosts for HIV-1, dengue virus, Leishmania and Mycobacterium tuberculosis. Yet macrophage research is hampered by lack of appropriate cell models for modelling infection by these human pathogens, because available myeloid cell lines are, by definition, not terminally differentiated like tissue macrophages. We describe here a method for deriving monocytes and macrophages from human Pluripotent Stem Cells which improves on previously published protocols in that it uses entirely defined, feeder- and serum-free culture conditions and produces very consistent, pure, high yields across both human Embryonic Stem Cell (hESC) and multiple human induced Pluripotent Stem Cell (hiPSC) lines over time periods of up to one year. Cumulatively, up to ∼3×10(7) monocytes can be harvested per 6-well plate. The monocytes produced are most closely similar to the major blood monocyte (CD14(+), CD16(low), CD163(+)). Differentiation with M-CSF produces macrophages that are highly phagocytic, HIV-1-infectable, and upon activation produce a pro-inflammatory cytokine profile similar to blood monocyte-derived macrophages. Macrophages are notoriously hard to genetically manipulate, as they recognise foreign nucleic acids; the lentivector system described here overcomes this, as pluripotent stem cells can be relatively simply genetically manipulated for efficient transgene expression in the differentiated cells, surmounting issues of transgene silencing. Overall, the method we describe here is an efficient, effective, scalable system for the reproducible production and genetic modification of human macrophages, facilitating the interrogation of human macrophage biology.
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iPSC-derived macrophages closely match tissue macrophages at both transcriptional and functional levels.
“Importantly, these cells closely match tissue macrophages at both transcriptional and functional levels (184–186).”
From Targeting macrophages in liver fibrosis (2026) · cites this paper as [185]
The Results section, “MDM Derived from PSC are Functionally Similar to b-MDM” (page unavailable), supports functional similarity to blood monocyte-derived macrophages. The Discussion (page unavailable) describes tissue-specific macrophage differentiation as future work; it does not establish that these cells closely match tissue macrophages transcriptionally or functionally. (LLM, conf=0.86)
Experimental — accuracy not yet measuredThe pMacpre yield is consistent with previously reported yields for this protocol, similar to Abud et al. 2017 (typically 40-fold), and 10-fold higher than two other recently published hiPS-microglia protocols.
“The cumulative yield of pMacpre for the lines used in this study was 10- to 43-fold higher than the number of input iPSCs, consistent with yields previously reported for this protocol (van Wilgenburg et al., 2013), similar to a recently published hiPS-microglia protocol (typically 40-fold) (Abud et al., 2017), and 10-fold higher than two other recently published hiPS-microglia protocols (0.5- to 4”
From A highly efficient human pluripotent stem cell microglia model displays a neuronal-co-culture-specific expression profile and inflammatory response (2017) · cites this paper as van Wilgenburg et al., 2013
The authors subsequently adapted the protocol to a fully defined, robust serum-free protocol.
“In 2008, we derived human PSC macrophages using a very simple methodology without OP9 feeders (Karlsson et al., 2008), which we subsequently adapted to a fully defined, robust serum-free protocol (van Wilgenburg et al., 2013), now widely used by others (Aflaki et al., 2014, Alasoo et al., 2015, Gupta et al., 2016).”
From A highly efficient human pluripotent stem cell microglia model displays a neuronal-co-culture-specific expression profile and inflammatory response (2017) · cites this paper as van Wilgenburg et al., 2013
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- Authors
- Wilgenburg, B. V., Browne, C., Vowles, J., & Cowley, S. A.
- Affiliations
- University of Oxford (GB)
- Corresponding author
- Cowley, Sally A.
- Venue
- PLoS ONE
- Publisher
- Public Library of Science (PLoS)
- Published
- 2013-08-12
- Volume
- 8
- Issue
- 8
- Pages
- e71098
- ISSN
- 1932-6203 (ISSN-L)
- PMC
- PMC3741356
- OpenAlex
- W2121999038
- Cited by
- 353
- References
- 44
- Citations by year
- 2026: 34 · 2025: 30 · 2024: 50 · 2023: 37 · 2022: 39 · 2021: 36 · 2020: 35 · 2019: 19 · 2018: 27 · 2017: 18 · 2016: 13 · 2015: 10 · 2014: 3 · 2013: 2
- FWCI
- 4.18
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS ONE (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, other-oa)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Funders
- Wellcome Trust (91070, WT082260, G090074791070, G0900747, LC0910-004, 090532, 090532/Z/09/Z, 090532/Z/09/, 090532/Z)
- Parkinson's UK (J-0901)
- Medical Research Council (G0900747 91070, Hub grant G0900747 91070, 090532/Z09/Z)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
- Health Sciences › Medicine › Hematology › Hematopoietic Stem Cell Transplantation
- Keywords
- monocyte-derived macrophages, human pluripotent stem cells, human embryonic stem cells, human induced pluripotent stem cells, defined culture conditions, serum-free culture, HIV-1 infection, lentiviral transduction, transgene expression, macrophage phagocytosis, inflammatory cytokines, macrophage infection model
- MeSH terms
- Cell Differentiation, Cell Line, Culture Media, Gene Expression Regulation, Humans, Macrophages, Monocytes, Phagocytosis, Phenotype, HIV-1, Macrophage Colony-Stimulating Factor, Cell Culture Techniques, Polymorphism, Single Nucleotide, Pluripotent Stem Cells, Embryonic Stem Cells, Induced Pluripotent Stem Cells
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