Figure 1.

2D Coculture of iNeurons and Pre-matured hAstros

(A) Example images showing the continued hPSC colony structure in the iNeuron line within 1 day after induction of NGN2, and the subsequent drastic change toward neuronal morphology by day 6. hAstros could be added to the iNeurons to be used for coculture studies.

(B) Cocultures of iNeurons (TUBB3-positive, nuclei are DAPI positive) with pre-matured hAstros (GFAP positive) are stable at 2 weeks. hAstro morphology becomes branched and elongated after 2 weeks of coculture, although not similar to that of astrocytes in the brain.

(C) Example of high-density neuronal fibers projecting out of iNeuron seeds (yellow arrows).

(D) hAstros (S100- and mGFP-positive) on high-density neuronal fibers induces morphological rearrangement that increases the number of branches and size compared with dissociated hAstros without iNeurons. n = 10 technical replicates for hAstros only, n = 11 for cocultures.

Error bars represent SEM. Scale bars are 50 μm.