Question about PC12
mdep at musica.mcgill.ca
Mon Jun 5 23:08:25 EST 2000
I use PC12 cells on collagen coated dishes. Although it is not conventinal,
I differentiate in 1-2% FBS in 50-100 ng/ml NGF (2.5S) and I change the
medium every 48 hours and do not let the cells go longer without feeding.
(this is essential). I had trouble with horse serum and could not get
consistently good cultures. I grow them on 60 mm dishes and I plate 1.5
million cells (avoid clumps when plating!!) and start differentiation 18-24
hours after plating.... don't wait more than a day to start
differentiation. These few points keep my cultures looking very good and I
get extensive neurites. Also, for my stock cultures, I change the medium
every 2 days and pass them (1:5 ratio) once a week for maintenace in 10% FBS
> I am doing some works on PC12E2 cells (a derivative of PC12 cells but
> attaches much better). they grow well in DMEM media with 10% FBS, 5%
> Horse serum, 1mM pyruvate, 10mM HEPES. but when i differentiated them by
> NGF (50 ng/ml) in DMEM with 10 mM HEPES, 0 or 1 or 2% horse serum, they
> first showed some kind of differentiation but after two or three days,
> they rounded up, formed clumps and finally, within 1 week, they all
> died. Does anyone have some idea or advise?
> thanks a lot!
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