Frank O. Fackelmayer
Frank.Fackelmayer at uni-konstanz.de
Mon Nov 22 03:39:44 EST 1999
"Ian A. York" wrote:
> I thought this was going to be an easy one to find in medline, but boy was
> I wrong.
> I want to induce necrotic cell death in several lines of cultured cells.
> I want this to be all necrosis, all the time; no apoptosis sneaking in
> here. And I want it to be a well-known protocol, so that we don't have to
> spend a lot of time justifying the system and running all the various
> controls to prove it's pure necrosis.
> But when I try to do searches, all I can find is people saying things
> like, "unlike necrotic cell death, Drug X induced apoptosis ... "
> I guess no one cares about boring old necrosis, but surely someone must
> have done a control some time.
> Any tips?
There ARE definitely ways to get pure necrosis in cell culture. One we used
with good success is to treat the cells with a combination of staurosporine
and oligomycin, as first described by Marcel Leist et al., 1997, J. Exp. Med.
185:1481-1486. In a paper from my group (Goehring et al. 1997; EMBO J.
16(24):7361-7371) that was done in cooperation with Marcel, we used this
system to demonstrate cleavage of a caspase substrate in apoptosis, but not
necrosis. The system works by killing cells with the kinase inhibitor
staurosporine (BTW, a great inducer of apoptosis in almost all cells) after
energy depletion. Without pretreatment with oligomycin, cells challanged with
staurosporine die via apoptosis, but in absence of intracellular ATP the mode
of cell death in completely necrotic. The system is easy to use and well
characterized. Drop me a line if you need more specific advice.
Hope this helps,
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