al., 2011), existing methods are slow (up to 6 months) (Jiang et al., 2013, Krencik et al., 2011, Shaltouki et al., 2013) and/or require sorting to reduce heterogeneity (Chaboub and Deneen, 2013, Yuan et al., 2011). Here, we screened a number of published protocols, along with commercially available media for primary human astrocyte culture, identifying a robust and straightforward differentiation protocol for generating astrocytes from hiPSCs. By co-culture with microglia, we compared the function of primary human fetal astrocytes and hiPSC-astrocytes in assays for neuroinflammatory response, phagocytosis, and spontaneous calcium activity, concluding that hiPSC-astrocytes are highly similar to their primary counterparts. Altogether, our rapid differentiation protocol, co-culture strategy, and scalable phenotypic assays will serve as a robust platform for queries of healthy and diseased human astrocytes.
