Human Pluripotent Stem Cell Differentiation to Microglia
Laraib Ijaz1, Madhura Nijsure1, Valentina Fossati2
1The New York Stem Cell Foundation Research Institute, New York, NY, USA.
Abstract:
Microglia, the immune cells of the central nervous system (CNS), play critical roles in CNS homeostasis and disease. Mounting evidence has linked aberrant microglial functions to neurodevelopment, neuroinflammatory and neurodegenerative diseases, underlining the need for novel models to investigate human microglia biology. Here we describe a protocol for generating in vitro patient-specific microglia progenitors and microglia-like cells from induced pluripotent stem cells (iPSCs). Our protocol generates microglia progenitor cells in approximately 35 days, which then can further mature into microglia-like cells within two additional weeks. Microglia differentiation is driven by specific growth factors and cytokines in serum-free conditions, resulting in mesodermal progenitors that grow in a monolayer which releases free-floating microglia progenitors. Isolated progenitors can be used in co-culture systems with other neuronal cells, xenotransplanted to generate chimeric mouse models, or further differentiated into adherent microglia-like cells for functional studies.
Insights
Researchers developed a new protocol using induced pluripotent stem cells (iPSCs) to create patient-specific microglia progenitors and microglia-like cells for studying central nervous system (CNS) diseases.
Area of Science:
- Neuroscience
- Immunology
- Stem Cell Biology
Background:
- Microglia are key immune cells in the central nervous system (CNS), crucial for homeostasis and implicated in various neurological disorders.
- Aberrant microglial function is linked to neurodevelopmental, neuroinflammatory, and neurodegenerative diseases.
- There is a significant need for advanced in vitro models to study human microglia biology.
Purpose of the Study:
- To establish a protocol for generating patient-specific microglia progenitors and microglia-like cells from induced pluripotent stem cells (iPSCs).
- To provide a novel in vitro model for investigating human microglia in health and disease.
Main Methods:
- Utilized induced pluripotent stem cells (iPSCs) for differentiation.
- Employed specific growth factors and cytokines in serum-free conditions.
- Generated mesodermal progenitors that release free-floating microglia progenitors.
Main Results:
- Successfully generated microglia progenitor cells within approximately 35 days.
- Achieved further maturation into microglia-like cells within an additional two weeks.
- Developed a protocol yielding adherent microglia-like cells for functional studies.
Conclusions:
- The protocol enables the generation of patient-specific microglia progenitors and microglia-like cells from iPSCs.
- These cells serve as valuable tools for co-culture systems, chimeric mouse models, and functional studies.
- This advancement facilitates research into CNS homeostasis and diseases involving microglial dysfunction.
