Related Experiment Video
Updated: Feb 27, 2026
Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
Published on: October 20, 2014
Modulation of Hematopoietic Lineage Specification Impacts TREM2 Expression in Microglia-Like Cells Derived From Human
Peter J Amos1, Susan Fung1, Amanda Case1
11 Department of Neurology, University of Washington, Seattle, WA, USA.
Abstract:
Microglia are the primary innate immune cell type in the brain, and their dysfunction has been linked to a variety of central nervous system disorders. Human microglia are extraordinarily difficult to obtain for experimental investigation, limiting our ability to study the impact of human genetic variants on microglia functions. Previous studies have reported that microglia-like cells can be derived from human monocytes or pluripotent stem cells. Here, we describe a reproducible relatively simple method for generating microglia-like cells by first deriving embryoid body mesoderm followed by exposure to microglia relevant cytokines. Our approach is based on recent studies demonstrating that microglia originate from primitive yolk sac mesoderm distinct from peripheral macrophages that arise during definitive hematopoiesis. We hypothesized that functional microglia could be derived from human stem cells by employing BMP-4 mesodermal specification followed by exposure to microglia-relevant cytokines, M-CSF, GM-CSF, IL-34, and TGF-β. Using immunofluorescence microscopy, flow cytometry, and reverse transcription polymerase chain reaction, we observed cells with microglia morphology expressing a repertoire of markers associated with microglia: Iba1, CX3CR1, CD11b, TREM2, HexB, and P2RY12. These microglia-like cells maintain myeloid functional phenotypes including Aβ peptide phagocytosis and induction of pro-inflammatory gene expression in response to lipopolysaccharide stimulation. Addition of small molecules BIO and SB431542, previously demonstrated to drive definitive hematopoiesis, resulted in decreased surface expression of TREM2. Together, these data suggest that mesodermal lineage specification followed by cytokine exposure produces microglia-like cells in vitro from human pluripotent stem cells and that this phenotype can be modulated by factors influencing hematopoietic lineage in vitro.
Insights
Researchers developed a simple method to generate human microglia-like cells from pluripotent stem cells. This advance aids studying brain disorders and the impact of genetic variants on microglia function.
Area of Science:
- Neuroscience
- Immunology
- Stem Cell Biology
Background:
- Microglia are crucial innate immune cells in the brain, implicated in various central nervous system disorders.
- Obtaining primary human microglia for research is challenging, hindering studies on genetic variants' impact.
- Previous methods for generating microglia-like cells include using human monocytes or pluripotent stem cells.
Purpose of the Study:
- To establish a reproducible and straightforward method for generating human microglia-like cells in vitro.
- To investigate the potential of deriving functional microglia from human pluripotent stem cells via mesodermal specification.
- To characterize the phenotype and function of these generated microglia-like cells.
Main Methods:
- Derived embryoid body mesoderm from human pluripotent stem cells.
- Exposed cells to microglia-relevant cytokines: M-CSF, GM-CSF, IL-34, and TGF-β.
- Utilized immunofluorescence microscopy, flow cytometry, and RT-PCR for cell characterization.
- Assessed functional phenotypes such as Aβ peptide phagocytosis and inflammatory gene expression.
Main Results:
- Successfully generated microglia-like cells exhibiting characteristic morphology and expressing key microglia markers (Iba1, CX3CR1, CD11b, TREM2, HexB, P2RY12).
- These cells demonstrated myeloid functional phenotypes, including phagocytosis of Aβ peptides and lipopolysaccharide-induced pro-inflammatory gene expression.
- Inhibition of definitive hematopoiesis (using BIO and SB431542) led to reduced TREM2 surface expression, suggesting modulation of the hematopoietic lineage.
Conclusions:
- Mesodermal lineage specification followed by specific cytokine exposure is an effective strategy for generating microglia-like cells in vitro from human pluripotent stem cells.
- This method provides a valuable tool for studying human microglia biology and its role in neurological diseases.
- The generated microglia-like cell phenotype can be influenced by factors affecting hematopoietic lineage development.
More Related Videos
05:35Transplantation of Human Induced Pluripotent Stem Cell-Derived Microglia in Immunocompetent Mice Brain via Non-Invasive Transnasal Route
Published on: May 31, 2022
10:25Pan-myeloid Differentiation of Human Cord Blood Derived CD34+ Hematopoietic Stem and Progenitor Cells
Published on: August 9, 2019
Related Concept Videos
01:21Lineage Commitment
01:01Regulation of Hematopoietic Stem Cells
01:19Multipotency of Hematopoietic Stem Cells
01:15Differentiation of Common Myeloid Progenitor Cells