Iscove’s Modified Dulbecco’s Medium

Protocol tips

Upstream tips	Protocol tips	Downstream tips
hESCs were dissociated to single cells and quickly reaggregated using low-cell-adhesion 96-well culture plate with V-bottomed wells (PrimeSurface MS-9096V; Sumitomo Bakelite)	cerebellar differentiation medium [Iscove’s modified Dulbecco’s medium/Ham’s F-12 1:1, 1 chemically defined lipid concentrate, penicillin/streptomycin, monothioglycerol (450 M) and purified BSA (99% purified by crystallization; Sigma) ](6,000 cells/well) containing growth-factor-free chemically defined medium (gfCDM), insulin (7 μg/ml), 10 μM Y-27632, and 10 μM SB431542 (Tocris Bioscience)

Upstream tips
hESCs were dissociated to single cells and quickly reaggregated using low-cell-adhesion 96-well culture plate with V-bottomed wells (PrimeSurface MS-9096V; Sumitomo Bakelite)
Protocol tips
cerebellar differentiation medium [Iscove’s modified Dulbecco’s medium/Ham’s F-12 1:1, 1 chemically defined lipid concentrate, penicillin/streptomycin, monothioglycerol (450 M) and purified BSA (99% purified by crystallization; Sigma) ](6,000 cells/well) containing growth-factor-free chemically defined medium (gfCDM), insulin (7 μg/ml), 10 μM Y-27632, and 10 μM SB431542 (Tocris Bioscience)

Publication protocol

cerebellar differentiation medium [ Iscove’s modified Dulbecco’s medium/Ham’s F-12 1:1, 1 chemically defined lipid concentrate, penicillin/streptomycin, monothioglycerol (450
M) and purified BSA (99% purified by crystallization; Sigma)] (6,000 cells/well) containing growth-factor-free chemically defined medium (gfCDM), insulin (7 μg/ml), 10 μM Y-27632, and 10 μM SB431542 (Tocris Bioscience)

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Iscove’s Modified Dulbecco’s Medium below.

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