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Updated: Dec 17, 2025
Transplantation of Human Induced Pluripotent Stem Cell-Derived Microglia in Immunocompetent Mice Brain via Non-Invasive Transnasal Route
Published on: May 31, 2022
Functional analysis of CX3CR1 in human induced pluripotent stem (iPS) cell-derived microglia-like cells
Nobuhito Murai1,2, Maisam Mitalipova1, Rudolf Jaenisch1,3
1Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
Abstract:
Microglia are the primary immune cells of the central nervous system and crucial to proper development and maintenance of the brain. Microglia have been recognized to be associated with neurodegenerative diseases and neuroinflammatory disorders. CX3C chemokine receptor 1 (CX3CR1), which is specifically expressed in microglia, regulates microglia homeostatic functions such as microglial activation and is downregulated in aged brain and disease-associated microglia in rodents, yet its role in human microglia is not fully understood. In this study, we investigated the function of CX3CR1 in human microglia using human induced pluripotent stem (iPS) cell-derived microglia-like cells. Human iPS cell-derived microglia-like cells expressed microglial markers and showed an activated state and phagocytic activity. Using CRISPR/Cas9 genome editing, we deleted CX3CR1 in human iPS cells and found increased inflammatory responses and phagocytic activity in mutant as compared to wild-type microglia-like cells. In addition, the CX3C chemokine ligand 1 (CX3CL1, a ligand for CX3CR1) significantly decreased the upregulation of IL-6 by lipopolysaccharide stimulation in human iPS cell-derived microglia-like cells. These results suggest that CX3CR1 in human microglia may contribute to microglial homeostasis by regulating inflammatory response and phagocytosis.
Insights
The CX3C chemokine receptor 1 (CX3CR1) plays a key role in human microglia function. Deleting CX3CR1 increased inflammatory responses and phagocytosis, suggesting its importance in brain immune cell homeostasis.
Area of Science:
- Neuroscience
- Immunology
- Stem Cell Biology
Background:
- Microglia are central nervous system immune cells vital for brain health.
- CX3C chemokine receptor 1 (CX3CR1) regulates microglial function but its role in human cells is unclear.
- CX3CR1 is downregulated in aged and diseased brains, suggesting a role in neurodegeneration.
Purpose of the Study:
- To investigate the function of CX3CR1 in human microglia.
- To understand CX3CR1's role in microglial inflammatory responses and phagocytosis.
- To utilize human induced pluripotent stem (iPS) cell-derived microglia-like cells for studying human microglia.
Main Methods:
- Generated human microglia-like cells from iPS cells.
- Utilized CRISPR/Cas9 genome editing to delete CX3CR1.
- Assessed inflammatory responses and phagocytic activity in CX3CR1-deficient and wild-type cells.
- Investigated the effect of CX3CL1 on lipopolysaccharide-induced IL-6 upregulation.
Main Results:
- Human iPS cell-derived microglia-like cells exhibited characteristic microglial markers, activation, and phagocytosis.
- Deletion of CX3CR1 led to heightened inflammatory responses and phagocytic activity compared to wild-type cells.
- CX3C chemokine ligand 1 (CX3CL1) reduced lipopolysaccharide-induced IL-6 increase in these cells.
Conclusions:
- CX3CR1 appears to regulate inflammatory responses and phagocytosis in human microglia.
- CX3CR1 may play a crucial role in maintaining microglial homeostasis in the human brain.
- These findings provide insights into the function of CX3CR1 in human neuroinflammation and disease.
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