With our astrocytes purchased from 3H biomedical we were advised to purchase a branded matrix coating 'Geltrex' and specific astrocyte media (AM). I was just wondering if anyone can advise on the benefits or otherwise of these, or which is the best generic media/coating for human astrocyte culture- the protocols i've read vary, with poly-l-lysine, collagen, and other coatings used and variations on DMEM and AM for the media.
for human astrocytes we use a basic MEM media supplemented with 10% serum, 1% pen/strep, 1% sodium pyruvate, 1% glutamine.
I coat my flasks with poly-L-lysine
For mouse, we use currently use regular tc dishes and DMEM/F12 (sensicell media from lifetechnologies A15155-01) supplemented with 10% FBS and anti-anti (antibiotic antibiotic). This isn't to say that the cells are "happier" on coated dishes versus uncoated, but it's what works when doing general cultures. If we want to image live cells though, we are currently growing on coverslips coated with 1 mg/ml Poly D Lysine (sigma).
We cultivate murine astrocytes (for immunofluorescence) on slides coated with poly-L-lysine in media based on RPMI 1640 with 10% FBS, 5% Pen/Strep and 5% glutamine.
Probably you resolved the problem because this question is old.....in other case...It is advisable to use matrix coating instead ply-L-Lysine; Geltrex or Matrigel ( there are many formulations, for example I am using Matrigel from BDsciences). Then, you can use different medium like the recipes here above. If you have to generate astrocytes from precursors, a very good medium is the NeuroCult™ NS-A Differentiation Kit (Human) from StemCell technologies.
We cultivate murine astrocytes coated with both poly-D-lysine and 0.2% gelatin in PBS. Culture astrocytes with 15% FBS, 1% Pen/Strep.
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