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After removal of the supernatant, the aggregates were resuspended in 3.5 ml/million cells of differentiation medium comprised of RPMI 1640 (Life Technologies) with B-27 either with or without insulin (RPMI/B-27 +/-) (Life Technologies) depending on the cell line and 3 or 4 M CHIR99021 (BOC Sciences)

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Makhro A, Hnggi P, Goede JS, Wang J, Brggemann A, Gassmann M, et al