To test drug application via an EC layer, type I collagen gel was poured into the left medium reservoir after injection of iALS-derived MN spheroids. The iECs (iCell ECs) were then seeded into the left well on the collagen gel (5.0 × 105 cells/ml). To accelerate differentiation into brain-specific ECs, RA (20 μM) was added (fig. S10A). After 7 days of coculture, drugs were applied to the left medium reservoir.

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