HL-1 cells were originally acquired from May Lam (mlam@lsuhsc.edu, 504-568-4737)
HL-1 medium
- 500 mL JRH Biosciences Claycomb medium
- 57.5 mL FBS
- 5.75 mL penicillin-streptomycin
- 5.75 mL norepinephrine solution (10 mM) (see below)
- 5.75 mL L-glutamine (200 mM)
Filter using a 0.2 μm Millipore Stericup filter into a 500 mL bottle
© Wrap the Claycomb Medium bottle in aluminum foil, since the medium is extremely light sensitive.
© Supplemented Claycomb medium is good for two weeks, at which time L-glutamine is replenished.
How to prepare a 10 mM norepinephrine solution?
Norepinephrine is made up in a 30 mM ascorbic acid solution
© Take 0.59 g ascorbic acid (Sigma) to 100 ml of distilled water
© Make sure all the ascorbic acid dissolves in the water → this is your 30 mM ascorbic acid solution
© Weigh out 320 mg norepinephrine (Sigma) and add 100 mL of the 30 mM ascorbic acid stock → this is your 10 mM norepinephrine solution
© Filter using a 0.2 μm Millipore Stericup filter into a 250 mL bottle
© Store the solution at -20°C
Note: Norepinephrine needs to be made up fresh on a monthly basis.