Help! neuronal cell culture problem

Serotone serotone at replies_to_newsgroup.com
Thu Aug 3 13:50:35 EST 2000


"Don Cates" <cates at cc.umanitoba.ca> wrote in message
news:39881d93.49487258 at news.cc.umanitoba.ca...
> 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?

Do these ropes consist of lysed cells or healthy ones?  Trypsin activity
remaining in the cells suspension would cause this.....

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