Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	DMEM/F12 supplemented with Chemically Defined Lipid Concentrate (1X) and N2-supplement (1X) and maintained in an incubator with 5% CO2 and 40% Oxygen. The aggregates were kept in this media for 15 to 20 days, at which time heparin (5 μM, Sigma-Aldrich, Natick, MA), FBS (10% v/v, Gemini Bio-Products, West Sacramento CA), and Matrigel (final 1% v/v, Corning Incorporated—Life Sciences, Oneonta, NY) were added to the medium.	On day 70 the amount of Matrigel was increased to 2% and B27 supplement (1X) was added to the medium.

Protocol tips
DMEM/F12 supplemented with Chemically Defined Lipid Concentrate (1X) and N2-supplement (1X) and maintained in an incubator with 5% CO2 and 40% Oxygen. The aggregates were kept in this media for 15 to 20 days, at which time heparin (5 μM, Sigma-Aldrich, Natick, MA), FBS (10% v/v, Gemini Bio-Products, West Sacramento CA), and Matrigel (final 1% v/v, Corning Incorporated—Life Sciences, Oneonta, NY) were added to the medium.
Downstream tips
On day 70 the amount of Matrigel was increased to 2% and B27 supplement (1X) was added to the medium.

Publication protocol

DMEM/F12 supplemented with Chemically Defined Lipid Concentrate (1X) and N2-supplement (1X) and maintained in an incubator with 5% CO2 and 40% Oxygen. The aggregates were kept in this media for 15 to 20 days, at which time heparin (5 μM, Sigma-Aldrich, Natick, MA), FBS (10% v/v, Gemini Bio-Products, West Sacramento CA), and Matrigel (final 1% v/v, Corning Incorporated—Life Sciences, Oneonta, NY) were added to the medium.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham below.

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