Gibco™DMEM/F-12

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	increasing amounts of N2 media (25, 50, 75, 100%) were added to the mTeSR medium every other day whilst maintaining 2.5uM dorsomorphin. N2 medium contains 1X N2 supplement (Invitrogen), 50% DMEM/F12, 50% Neurobasal, 0.075% Bovine serum albumin, and 2mM GlutaMAX (Life Technologies)	Cells were subsequently passaged with trypsin and cultured on matrigel-coated dishes in N2 media with the addition of 20ng/ml bFGF. Rho-associated protein kinase (ROCK) inhibitor Y27632 (10uM, Stemgent) was added to the medium for the first 24 hours. NPs were expanded and maintained in N2 medium with 20ng/ml bFGF. All 2D culture experiments were performed on WIBR3-derived NPs unless otherwise specified.

Protocol tips
increasing amounts of N2 media (25, 50, 75, 100%) were added to the mTeSR medium every other day whilst maintaining 2.5uM dorsomorphin. N2 medium contains 1X N2 supplement (Invitrogen), 50% DMEM/F12, 50% Neurobasal, 0.075% Bovine serum albumin, and 2mM GlutaMAX (Life Technologies)
Downstream tips
Cells were subsequently passaged with trypsin and cultured on matrigel-coated dishes in N2 media with the addition of 20ng/ml bFGF. Rho-associated protein kinase (ROCK) inhibitor Y27632 (10uM, Stemgent) was added to the medium for the first 24 hours. NPs were expanded and maintained in N2 medium with 20ng/ml bFGF. All 2D culture experiments were performed on WIBR3-derived NPs unless otherwise specified.

Publication protocol

increasing amounts of N2 media (25, 50, 75, 100%) were added to the mTeSR medium every other day whilst maintaining 2.5uM dorsomorphin. N2 medium contains 1X N2 supplement (Invitrogen), 50% DMEM/F12, 50% Neurobasal, 0.075% Bovine serum albumin, and 2mM GlutaMAX (Life Technologies)

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Manufacturer protocol

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