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Microglia-containing neural organoids as brain microphysiological systems for long-term culture

Alex Rittenhouse1, Caroline Krall1,2, Jesse Plotkin1

  • 1Center for Alternatives to Animal Testing, Department of Environmental Health and Engineering, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD, United States.

Frontiers in Cellular Neuroscience
|October 20, 2025
PubMed

Insights

Researchers developed a novel microglia-integrated brain organoid model (μbMPS) for studying neurodevelopment and neuroinflammation. This scalable model successfully incorporates microglia, enhancing neuronal activity and maturity for research applications.

Area of Science:

  • Neuroscience
  • Developmental Biology
  • Immunology

Background:

  • Microglia are crucial for brain development, homeostasis, and neuroinflammation.
  • Standard brain organoid models often lack microglia due to their embryonic yolk sac origin.
  • Incorporating microglia is essential for accurate modeling of neurodevelopmental and neuroinflammatory processes.

Purpose of the Study:

  • To develop a reproducible microglia-integrated brain organoid model.
  • To assess the long-term survival, maturation, and functionality of integrated microglia.
  • To evaluate the impact of microglia on neuronal activity and maturity in organoids.

Main Methods:

  • Aggregation of hiPSC-derived neural and microglia progenitors in U-bottom 96-well plates.
  • Development of a microglia-integrated brain microphysiological system (μbMPS).
  • Long-term culture (over 9 weeks) of organoids with integrated microglia.

Main Results:

  • Successfully integrated, matured, and long-term survival of microglia within the neural organoid environment.
  • Demonstrated functional microglia activity, including phagocytosis and neuroinflammatory responses.
  • Observed enhanced neuronal activity and maturity in microglia-containing organoids.
  • The μbMPS model showed scalability and reproducibility.

Conclusions:

  • The microglia-integrated brain organoid model (μbMPS) provides a robust platform for studying brain development and function.
  • This model enables investigation of neuroinflammation and neurodevelopmental disorders.
  • The μbMPS is a valuable tool for neurotoxicology research and drug discovery.
Keywords:
iPSC-derived 3D modelsmicrogliamicrophysiological systems (MPS)neural organoidsnew approach methodologies (NAMs)

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