Development Of Organoid Based Model To Study Immune-Neural Interactions In Human Cns Diseases

Modeling neural-immune interactions in neurodegenerative and immune-mediated central nervous system (CNS) diseases requires human 3D models that capture cellular diversity and long-term tissue maturation. Here, we present an enhanced human induced pluripotent stem cell (hiPSC)-derived cerebral organoid (CO) platform optimized to mitigate core hypoxia for over 200 days. Timed pro-myelinating cues established organized neuronal layering and progressive axonal myelination through day 140, while vascular fusion yielded assembloids incorporating endothelial structures and microglia. Extended culture (>500-750 days) spontaneously reproduced hallmark features of human CNS aging, including cellular senescence signatures, neuroaxonal loss, hypomyelination, and the autonomous emergence of a neurotoxic astrocyte transcriptional profile in the complete absence of microglia or immune cells. Co-culture with autologous activated peripheral blood mononuclear cells (PBMC) resulted in transient immune infiltration and a pronounced type II interferon response across CNS lineages. High-plex spatial transcriptomics revealed that immune cell infiltration was associated with oligodendrocyte loss and in aged organoids also with downregulated oligodendrocyte myelin gene transcription. While not fully reproducing adult tissue stoichiometry, this platform enables longitudinal modeling of neural-immune crosstalk in age-related and neuroinflammatory CNS disorders.