Help! neuronal cell culture problem

Don Cates cates at
Wed Aug 2 08:21:05 EST 2000

I've been doing cell culture of dissociated neurons from fetal rat
medulla for several years. Recently the wheels have fallen off.
I use 35 mm plastic dishes with a collagen (Vitrogen) coating. I then
plate with neuron free medullary background cells for a week or more,
then plate with my dissociated cells (mince with scissors, treat with
trypsin for 15-20 min, gently triturate). The last few times I have done
this, the cells seem to lift off most of the dish and form long, thick
'ropes' of cells which include the collagen and background cells. I've
replaced my various media and components with fresh material with no
luck. It is not invariable, I get the odd good dish.
Can anyone suggest anything?

Don Cates
cates at

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