To more fully characterize the release of inflammatory mediators from control hiPSC-astrocytes following 24-hr treatment with 5 μM Aβ42, a main component of the amyloid plaques in AD, we measured proinflammatory (IL-1β, IL-4, IL-6, IL-8, IL-10, and tumor necrosis factor α) and anti-inflammatory cytokines (IL-1α, IL-2, IL-12, IL-17α, interferon-γ, and granulocyte macrophage colony-stimulating factor) by Multi-Analyte ELISArray, confirming that Aβ42 treatment primarily increased IL-6 secretion (Garwood et al., 2011), as well as a second proinflammatory cytokine, IL-8 (Figures S3B and S3C). We also measured 36 cytokines, chemokines, and acute-phase proteins using the Proteome Profiler Human Cytokine Array (Table S5) in baseline conditions and following 24-hr treatment with 5 μM Aβ42 (Figures 3D and S3C–S3E); Aβ42 increased IL-6 release in both hiPSC-astrocytes and primary human fetal astrocytes (Figure S3D). Together, our findings indicate that hiPSC-astrocytes are capable of secreting proinflammatory cytokines in response to neuroinflammatory cues.
