2. Academic Validation
  3. Protocol to generate microglia-containing cerebral organoids to model HIV neuroinflammation

Protocol to generate microglia-containing cerebral organoids to model HIV neuroinflammation

  • STAR Protoc. 2026 Jun 19;7(2):104604. doi: 10.1016/j.xpro.2026.104604.
Janet P Zayas 1,  David Gagliardi 1,  Stephanie Gambut 1,  Sam Acors 2,  Nathalia Almeida 2,  Michael H Malim 2,  Luis Apolonia 3,  Lena Al-Harthi 1,  Srinivas D Narasipura 4,  João I Mamede 5
Affiliations

Affiliations

  • 1 Department of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, IL 60612, USA.
  • 2 Department of Infectious Diseases, School of Immunology and Microbial Sciences, King's College London, London SE1 9RT, England.
  • 3 Department of Infectious Diseases, School of Immunology and Microbial Sciences, King's College London, London SE1 9RT, England; Centre for Gene Therapy & Regenerative Medicine, School of Basic & Medical Biosciences, King's College London, London SE1 9RT, England.
  • 4 Department of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, IL 60612, USA. Electronic address: [email protected].
  • 5 Department of Microbial Pathogens and Immunity, Rush University Medical Center, Chicago, IL 60612, USA. Electronic address: [email protected].
PMID: 42224077 DOI: 10.1016/j.xpro.2026.104604
Abstract

Cerebral organoids (COs) are valuable for studying neurodegenerative diseases and pathogens, but their limited microglia pose challenges. Here, we present a protocol to generate organoids with physiologically relevant microglia (CO-iMs). We describe steps for differentiating induced pluripotent stem cells (iPSCs) into mesoderm, seeding embryoid bodies (EBs), and harvesting hematopoietic progenitor cells (HPCs). We detail procedures for co-culturing HPCs and iPSCs to generate CO-iMs, followed by their Infection and treatment. The model provides a platform to study viral infections and neuroinflammation. For complete details on the use and execution of this protocol, please refer to Narasipura et al.1.

Keywords

Biotechnology and bioengineering; Microbiology; Organoids; Stem Cells.

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