N-2 MAX Media Supplement: R&D Systems

N-2 MAX Media Supplement (Catalog # AR009) is a serum-free, chemically defined, concentrated media supplement formulated to provide optimal growth conditions for neural stem cell expansion. N-2 MAX is composed of Recombinant Human Insulin, Human Transferrin, Putrescine, Selenite, and Progesterone. The supplement is supplied as a 100X concentrate in water.

Components

Recombinant Human Insulin (2500 µg/mL)
Human Transferrin (10,000 µg/mL)
Putrescine (1611 µg/mL)
Selenite (0.52 µg/mL)
Progesterone (0.63 µg/mL)

N-2 MAX Medium Preparation

Option 1: Mix the following components with deionized or distilled water to make 500 mL of medium. Adjust the pH to 7.2. Filter the solution (2 μm filter unit) and add 5 mL of 100X sterile Penicillin-Streptomycin solution (Invitrogen^®, Catalog # 15140-148). The medium may be stored in the dark at 2 °C to 8 °C for up to 2 weeks.

DMEM/F-12 (Invitrogen, Catalog # 12500-062) 6 g
Glucose (Sigma, Catalog # G6152) 0.775 g
Glutamine (Sigma, Catalog # G8540) 0.0365 g
NaHCO₃ (Sigma, Catalog # S5761) 0.845 g
N-2 MAX Media Supplement (100X) 5 mL

Option 2: Dilute 100-fold with a basal media (e.g. Neurobasal Media from ThermoFisher Scientific, Catalog # 21103-049) before use. The medium may be stored in the dark at 2 °C to 8 °C for up to 2 weeks.

Data Examples

Expansion and Differentiation of Rat Neural Stem Cells

A.		Culturing Neural Cell Types in Media Supplemented with N-2 MAX. Rat Cortical Stem Cells (Catalog # NSC001) were grown and differentiated for 7 days in vitro in media supplemented with N-2 MAX Media Supplement (Catalog # AR009). (A) beta-III Tubulin was detected in differentiated neurons using a neuron-specific Mouse beta-III Tubulin Monoclonal (clone TuJ-1) Antibody (Catalog # MAB1195), followed by NorthernLights™ 557-conjugated Donkey Anti-Mouse Secondary Antibody (Catalog # NL007; yellow). (B) GFAP was detected in differentiated astrocytes using a Sheep Anti-Human GFAP Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2594), followed by NorthernLights™ 557-conjugated Donkey Anti-Sheep Secondary Antibody (Catalog # NL010; yellow). (C) O4 was detected in differentiated oligodendrocytes using a Mouse Anti-Human/Mouse/Rat/Chicken Oligodendrocyte Marker O4 Monoclonal Antibody (Catalog # MAB1326), followed by NorthernLights 557-conjugated Goat Anti-Mouse Secondary Antibody (Catalog # NL019; yellow). The nuclei were counterstained with DAPI (blue).
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