[Histonet] Vasculature in tumors - long response

Tue Jul 5 18:24:10 CDT 2011

I also agree with Liz's well said synopsis. Be aware that in mouse and human CD34 is not as "pan" EC as CD31 or VE-cadherin, it is very endothelial bed selective.

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-----Original Message-----
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Sender: histonet-bounces <@t> lists.utsouthwestern.edu
Date: Tue, 5 Jul 2011 13:15:00 
To: 'Elizabeth Chlipala'<liz <@t> premierlab.com>; 'Leiker, Merced'<leiker <@t> buffalo.edu>; 'JR R'<rosenfeldtek <@t> hotmail.com>; <histonet <@t> lists.utsouthwestern.edu>
Subject: RE: [Histonet] Vasculature in tumors - long response

I agree with Liz on most all of this about CD31 and love the Dianova rat
anti ms ab on mouse xenographs, I have also tried the BC rat < ms anti CD31
and it works but not as good as Dianova for the really early vessels.
Another alternative I have used in the past on rats besides F8 and SMA is SC
goat < CD34 but the problem with CD34 is that it stains lots of other things
besides endothelial cells, it stains all progenitor cells, but that has been
an option for rat tissue.  Always looking for good endothelial cell markers
for rat especially but all species.

Regards,
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
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Aurora, CO 80010
P-720-859-4060
F-720-859-4110
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Elizabeth
Chlipala
Sent: Tuesday, July 05, 2011 11:10 AM
To: 'Leiker, Merced'; 'JR R'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Vasculature in tumors - long response

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
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com

Ship to address:

1567 Skyway Drive, Unit E
Longmont, CO 80504

-----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

> Date: Fri, 1 Jul 2011 12:48:33 -0500
> From: sgoebel <@t> mirnarx.com
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Vasculature in tumors
>
> Hello all and happy long weekend (yes our boss gave us the rest of the
> day off at 2!!!)
>
>
>
> Looking to stain vasculature in tumors.  I did trichrome, pentachrome,
> and PAS (with digestion) and am being told that these are not optimal
> for what they are looking for.  Any other ideas to stain blood vessels
> would be awesome!!
>
>
>
> Thanks ya'll!
>
>
>
> Sarah Goebel-Dysart, BA, HT(ASCP)
>
> Histotechnologist
>
> Mirna Therapeutics
>
> 2150 Woodward Street
>
> Suite 100
>
> Austin, Texas  78744
>
> (512)901-0900 ext. 6912
>
>
>
> _______________________________________________
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