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Updated: Nov 22, 2025
Transplantation of Human Induced Pluripotent Stem Cell-Derived Microglia in Immunocompetent Mice Brain via Non-Invasive Transnasal Route
Published on: May 31, 2022
Stem-cell-derived human microglia transplanted into mouse brain to study human disease
Nicola Fattorelli1,2, Anna Martinez-Muriana1,2, Leen Wolfs1,2
1Centre for Brain and Disease Research, Flanders Institute for Biotechnology (VIB), Leuven, Belgium.
Abstract:
Microglia are critically involved in complex neurological disorders with a strong genetic component, such as Alzheimer's disease, Parkinson's disease and frontotemporal dementia. Although mouse microglia can recapitulate aspects of human microglia physiology, they do not fully capture the human genetic aspects of disease and do not reproduce all human cell states. Primary cultures of human microglia or microglia derived from human induced pluripotent stem cells (PSCs) are difficult to maintain in brain-relevant cell states in vitro. Here we describe MIGRATE (microglia in vitro generation refined for advanced transplantation experiments, which provides a combined in vitro differentiation and in vivo xenotransplantation protocol to study human microglia in the context of the mouse brain. This article details an accurate, step-by-step workflow that includes in vitro microglia differentiation from human PSCs, transplantation into the mouse brain and quantitative analysis of engraftment. Compared to current differentiation and xenotransplantation protocols, we present an optimized, faster and more efficient approach that yields up to 80% chimerism. To quantitatively assess engraftment efficiency by flow cytometry, access to specialized flow cytometry is required. Alternatively, the percentage of chimerism can be estimated by standard immunohistochemical analysis. The MIGRATE protocol takes ~40 d to complete, from culturing PSCs to engraftment efficiency assessment.
Insights
Researchers developed MIGRATE, a novel protocol for generating human microglia from stem cells for neurological disease research. This method enhances the study of human microglia in mouse models, improving disease insights.
Area of Science:
- Neuroscience
- Stem Cell Biology
- Immunology
Background:
- Microglia play a crucial role in neurological disorders like Alzheimer's and Parkinson's disease.
- Existing methods for studying human microglia in mouse models have limitations in capturing human genetic disease aspects and cell states.
- Human microglia and stem cell-derived microglia are challenging to maintain in brain-relevant states in vitro.
Purpose of the Study:
- To present MIGRATE, a combined in vitro differentiation and in vivo xenotransplantation protocol for studying human microglia in the mouse brain.
- To provide a detailed, step-by-step workflow for human microglia generation, transplantation, and engraftment analysis.
- To offer an optimized, faster, and more efficient alternative to current protocols for human microglia xenotransplantation.
Main Methods:
- Human induced pluripotent stem cells (PSCs) differentiation into microglia.
- Xenotransplantation of differentiated human microglia into the mouse brain.
- Quantitative analysis of microglia engraftment using flow cytometry or immunohistochemistry.
Main Results:
- The MIGRATE protocol enables the study of human microglia within the mouse brain microenvironment.
- Achieved up to 80% chimerism, indicating high engraftment efficiency.
- Provides a faster and more efficient approach compared to existing protocols, with a total duration of approximately 40 days.
Conclusions:
- MIGRATE offers a robust and efficient method for generating and xenotransplanting human microglia.
- This protocol facilitates advanced research into human microglia function in neurological diseases within a relevant in vivo context.
- The improved efficiency and speed of MIGRATE advance the study of human microglia in complex neurological disorders.
