[BioC] OT: cell lines and tissues

Dapeng Cui bioddc at langate.gsu.edu
Wed Jul 7 14:57:56 CEST 2004


My suggestion is to do microdissection and amplify both tissues starting
from the same amount of RNA/cells.

Dapeng Cui

>>> Stefano Calza <stecalza at tiscali.it> 07/07/04 08:02AM >>>
Hi Rohit,

this was my point. But their (and mine at the statistical analysis
step)  problem is that from the normal tissue itself they cannot collect
enough RNA. Then, what could be the best (or least bad) 
solution? 
Microdissection would bring into the normal sample a mixture of cells,
not only epithelial ones. Amplifying RNA from the normal tissue (they
already have the chips for th patholigical ones) would 
probably bias the expression, though I guess the overall expression (or
also differentially at gene level?).

Summarizing: what could be the best procedure?

TIA,
Stefano

On Tue, Jul 06, 2004 at 08:18:13PM +0200, Ghai, Rohit wrote:
> hi Stefan
> 
> I feel that such comparisons would not offer as much contrast 
> and clarity. Normal tissues would be best, even if isolation
> is tedious. Cell lines grow in a completely different environment
> than tissues. Gene expression is critically dependent on outside
> cues from other cells in the tissue too. Of course, this also depends
on
> what is the question you are asking. If its a question of
identifying
> markers, these may be identified as one can then verify individual
markers
> on a smaller scale. But for a better distinction of the processes
underlying
> the disease state it would be better to use normal ovarian
epithelium.
> 
> regards
> Rohit 
> 
>  ------------------------------------------------------------------
> Hi everybody. 
> 
> This is definitely Off-Topic, but I'd like to have an opinion from
the many
> biologist (but not only!) that populate the list, about the
following
> problem:
> 
> A group of biologists is willing to study gene expression in ovarian
cancer
> tissues relative to normal ones. As the normal ovarian epithelium is
single
> layer, it's quite hard to get enough RNA. So 
> 
> they are actually going to compare normal ovarian cell lines grown in
vitro
> versus patological tissues. I feel a bit confused about this.
Wouldn't be
> better to amplify the RNA from normal 
> 
> tissues? Any other options? 
> 
> Any insight will be very appreciated. 
> 
> TIA, 
> Stefano
> 
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