[Histonet] how to count nodules in a xenograft model

[Liz Chlipala]

Caroline

I have done this in the past but with a different cell line B16F10Mel it's a
common animal model of metastatic melanoma.  The route is the same iv
injection and then counting the number of lesions in the lung.  The first
thing is at necropsy is to inflate the lungs with fixative, if she has not
done this before I believe there are other posts addressing this on the
Histonet or even possibly the web, but if she needs some info on that just
have her personally e-mail me and I'll go over the protocol.  From there we
did two things we counted the number of tumors on the outside of the lung
first, this can be easy or hard depending upon the size of the tumors,
sometimes there are too many to count.  From there we would process the lung
for routine H&E.  This can be done two ways; 1.  process the entire lung
whole and embed as flat as possible or 2.  Remove each of the lobes and then
process (if you choose to separate the lobes in processing it might not be a
bad idea to count the tumors on the outside of the lung for each lobe, that
way you could see if your necropsy data correlates with the histology data).

I find when the lobes are separated it's a bit easier to count the number of
tumors.  There should be some publications on this, for the B16F10Mel cell
line, I would search that on the web. I would initially start by just
cutting one section and staining with H&E and counting.  If she is comparing
treatment groups and there are significant differences between the
treatments she should be able to see differences by counting just one level.
If she is starting to see a trend and maybe the possibility that there are
differences in the treatment groups she could then make the decision to cut
additional levels and count those also.  I've only use the MDA-MB-231 cell
line as a sq xenograft before so I'm not certain how visible the tumors
would be in the lung sections.  It is a breast cancer cell line so some
immunostain might be helpful possibly keratin?  But why do the immunostain
if you can see the tumors in the H&E.  If she has any questions she can
contact me directly.  Hope this helps.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
 
Ship to Address:
 
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Caroline
Bass
Sent: Friday, October 06, 2006 9:51 PM
To: HISTONET
Subject: [Histonet] how to count nodules in a xenograft model

Hello Everyone,

I am posting this on behalf of a friend.  I know very little about  
xenograft models and I thought someone on this list may have more  
experience.

Essentially, she is injecting MDA-MB-231 cells i.v. in mouse and  
studying metastasis.  She wants to get a count of the nodules that  
form in the lung.  Does anyone have a good protocol for doing this.   
She has little histology skills, so most likely she will send the  
tissue to a lab to process.  What is the safest way to collect the  
tissue, how should it be processed, and how does one get a good idea  
of the number of nodules formed?

Any help would be appreciated.

Thanks,

Caroline


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