[Histonet] Vasculature in tumors

[Troutman, Kenneth A]

We use CD34 for endothelial staining in blood vessels and Podoplanin (D2-40) for lymphatic vessels.  These are for humans only.  (Mine was worth less than a half-cent...but hopefully helpful.)  :)

Ashley Troutman BS, HT(ASCP) QIHC
Immunohistochemistry Supervisor
Vanderbilt University Histopathology
1301 Medical Center Drive TVC 4531
Nashville, TN  37232

Date: Tue, 5 Jul 2011 11:10:02 -0600
From: Elizabeth Chlipala <liz <@t> premierlab.com>
Subject: RE: [Histonet] Vasculature in tumors - long response
To: "'Leiker, Merced'" <leiker <@t> buffalo.edu>, 'JR R'
      <rosenfeldtek <@t> hotmail.com>, "histonet <@t> lists.utsouthwestern.edu"
      <histonet <@t> lists.utsouthwestern.edu>
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      <14E2C6176416974295479C64A11CB9AE0646F756 <@t> SBS2K8.premierlab.local>
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Hello all

I have been following this discussion and I'm going to put in my 2 cents.  Tumor vasculature has traditionally been measured with IHC and CD31 or possibly another IHC marker.

We use CD31 here for mouse xenograft samples.  We have several antibodies we use and it is dependent upon the species.  Human tumors (not xenografts) are stained with CD31 but a different antibody (Dako) than the mouse xenografts (Dianova).  For other species, canine, porcine, etc we use factor VIII.  Factor VIII is not the best marker since it may not pick up fine capillaries but we have been unsuccessful in getting a CD31 to work in FFPE tissue for those species and we have tried several antibodies.  The dako antibody will also work on rabbit samples.  I can't remember off the top of my head on what we use for rat but it may be Factor VIII.

I believe that a large number of publications in which tumor vasculature is measured CD31 has been used.  I do not believe that any special stain will provide you with the information that is needed for this type of staining and analysis, especially if you are going to publish the results.

The IHC stain for CD31 (for mouse xenograft samples) had been tricky in the past since most antibodies that stain mouse endothelial cells only worked in frozen sections or zinc fixed (not zinc formalin) paraffin embedded samples.  Santa Cruz did have a goat polyclonal antibody that did work on FFPE samples for a period of time, early 2000's, but that antibody no longer works in FFPE samples (only frozen, something about the goat dying and the new lots of antisera did not work in FFPE tissues).  If you are staining mouse xenograft samples then the Dianova antibody would be the best one to use in my opinion.  I need to state that we have not used or even tried VE Cadherin or other IHC markers for vessels, so I really can't comment on those, they may work very well.

We use CD31 since we are also running analysis and in order to do that you need a very clean stain and we can achieve that with the Dianova antibody, prior to the Dianova antibody we had used the santa cruz antibody (we had the particular lot number that worked in FFPE tissues, which also worked in multiple species, I do miss that antibody).

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Leiker, Merced
Sent: Tuesday, July 05, 2011 10:39 AM
To: 'JR R'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Vasculature in tumors

I agree... I've used CD31 to stain tumor vasculature. Some tumor vasculature isn't even lined with endothelium; there's just blood "channels." But most should be lined.

Regards,
Merced

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of JR R
Sent: Friday, July 01, 2011 1:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Vasculature in tumors



You are probably looking for capillaries maybe arterioles so there wont be any elastic lamina for the pentachrome stain to detect.

Instead try IHC with an endothelial marker like VE Cadherin (best) or in a pinch, CD31 or PECAM.

Jerry Ricks
Research Scientist
University of Washington
Department of Pathology