EXTERNAL] Shipping spleens

Yes, I did this all the time.
Try this and see if you get better results.
Dissociate the spleen and don't put on ice - leave at Room temperature in serum
containing media (50ml) - you'll need to wash before fusion of course.
I think it works best if you don't lyse the cells until you are ready to use
them.
Hope this helps
Alan

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The Jackson Laboratory
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<email obscured>
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Louise Lopez
Office Administrator, Scientific Services
The Jackson Laboratory
600 Main Street, Bar Harbor, Maine 04609
P:  207-288-6484 ~ F:  207-288-6044 ~ C:  207-266-1419

The Jackson Laboratory: Leading the search for tomorrow’s cures
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-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Frances
Weis-Garcia <email obscured>>
Reply-To: <email obscured>" <email obscured>>
Date: Friday, April 23, 2021 at 1:13 PM
To: <email obscured>" <email obscured>>
Subject: [EXTERNAL][ABRF Ab Forum] Shipping spleens

    Does anyone have experience shipping spleens?  Good or bad?

    A researcher has immunized mice and would like to ship me the spleen for me
to prep cells from and then fuse them.  We did a test run ... harvested the
spleen yesterday ... put it in a tube of media ... let it sit on ice packs in a
box overnight ... prepared a single cell suspension in parallel to a freshly
harvested spleen.  We recovered only 33% of the cells, relative to the freshly
prepared ones.  Lots of gooey tissue left behind after squeezing it between 2
frosted slides but the cells were 74% viable (Fresh were 94%).

    I have never had good success with frozen splenocytes.

    Thoughts?
    ――
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Hi Kathy,
when we did the 4C wet ice shipping we found that the white cells stuck
together and we couldn't get them to dissociate. But I agree with Frances, best
to try both methods and see.
One other possibility is to get them into >34M Sucrose on ce which is what we
used to do when making macrophage feeder layers back in the day.... (God I'm
old!)
All the best
Alan

Rest of post

--
If you have used any of JAX’s  Scientific Services please remember to
acknowledge the Service(s) used, in all manuscripts, publications, grant
applications, press releases, and presentations.

Sample language: We gratefully acknowledge the contribution of [optional: name
of person] and the [insert Service’s name] Service at The Jackson Laboratory
for expert assistance with the work described in this publication.

Alan Sawyer
Senior Director, Scientific Services
The Jackson Laboratory
600 Main Street
Bar Harbor, ME 04609
Home Office: 207-276-8025
<email obscured>
www.jax.org
https://orcid.org/0000-0001-5942-4498

Louise Lopez
Office Administrator, Scientific Services
The Jackson Laboratory
600 Main Street, Bar Harbor, Maine 04609
P:  207-288-6484 ~ F:  207-288-6044 ~ C:  207-266-1419

The Jackson Laboratory: Leading the search for tomorrow’s cures
--
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-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of
"Brundage, Kathleen" <email obscured>>
Reply-To: <email obscured>" <email obscured>>
Date: Friday, April 23, 2021 at 2:10 PM
To: <email obscured>" <email obscured>>
Subject: [EXTERNAL]Re: [ABRF Ab Forum] Shipping spleens

    Frances,
     I suggest you make a single cell suspension of the spleen put it in a 15ml
screw cap tube and fill it with media + 10% FCS/FBS. Make sure there a minimal
air pocket and ship it on wet ice.
    I know if we put single cell suspension of spleens in media in an ice
bucket in the fridge overnight the cell viable does not change drastically.
One other thing, this is usually after rbc lysis.
    Kathy

    ________________________________
    From: Antibody Discussion Forum <email obscured>> on behalf of
Frances Weis-Garcia <email obscured>>
    Sent: Friday, April 23, 2021 1:12 PM
    To: <email obscured> <email obscured>>
    Subject: [ABRF Ab Forum] Shipping spleens

    Does anyone have experience shipping spleens?  Good or bad?

    A researcher has immunized mice and would like to ship me the spleen for me
to prep cells from and then fuse them.  We did a test run ... harvested the
spleen yesterday ... put it in a tube of media ... let it sit on ice packs in a
box overnight ... prepared a single cell suspension in parallel to a freshly
harvested spleen.  We recovered only 33% of the cells, relative to the freshly
prepared ones.  Lots of gooey tissue left behind after squeezing it between 2
frosted slides but the cells were 74% viable (Fresh were 94%).

    I have never had good success with frozen splenocytes.

    Thoughts?
    ――
    View topic
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that should be 0.34M not 34M

Rest of post

--
If you have used any of JAX’s  Scientific Services please remember to
acknowledge the Service(s) used, in all manuscripts, publications, grant
applications, press releases, and presentations.

Sample language: We gratefully acknowledge the contribution of [optional: name
of person] and the [insert Service’s name] Service at The Jackson Laboratory
for expert assistance with the work described in this publication.

Alan Sawyer
Senior Director, Scientific Services
The Jackson Laboratory
600 Main Street
Bar Harbor, ME 04609
Home Office: 207-276-8025
<email obscured>
www.jax.org
https://orcid.org/0000-0001-5942-4498

Louise Lopez
Office Administrator, Scientific Services
The Jackson Laboratory
600 Main Street, Bar Harbor, Maine 04609
P:  207-288-6484 ~ F:  207-288-6044 ~ C:  207-266-1419

The Jackson Laboratory: Leading the search for tomorrow’s cures
--
--



-----Original Message-----
From: Antibody Discussion Forum <email obscured>> on behalf of Alan
Sawyer <email obscured>>
Reply-To: <email obscured>" <email obscured>>
Date: Friday, April 23, 2021 at 3:22 PM
To: <email obscured>" <email obscured>>
Subject: Re: [ABRF Ab Forum] [EXTERNAL]Re: Shipping spleens

    Hi Kathy,
    when we did the 4C wet ice shipping we found that the white cells stuck
together and we couldn't get them to dissociate. But I agree with Frances, best
to try both methods and see.
    One other possibility is to get them into >34M Sucrose on ce which is what
we used to do when making macrophage feeder layers back in the day.... (God I'm
old!)
    All the best
    Alan
    --
    If you have used any of JAX’s  Scientific Services please remember to
acknowledge the Service(s) used, in all manuscripts, publications, grant
applications, press releases, and presentations.

    Sample language: We gratefully acknowledge the contribution of [optional:
name of person] and the [insert Service’s name] Service at The Jackson
Laboratory for expert assistance with the work described in this publication.

    Alan Sawyer
    Senior Director, Scientific Services
    The Jackson Laboratory
    600 Main Street
    Bar Harbor, ME 04609
    Home Office: 207-276-8025
    <email obscured>
    www.jax.org
    https://orcid.org/0000-0001-5942-4498

    Louise Lopez
    Office Administrator, Scientific Services
    The Jackson Laboratory
    600 Main Street, Bar Harbor, Maine 04609
    P:  207-288-6484 ~ F:  207-288-6044 ~ C:  207-266-1419

    The Jackson Laboratory: Leading the search for tomorrow’s cures
    --
    --



    -----Original Message-----
    From: Antibody Discussion Forum <email obscured>> on behalf of
"Brundage, Kathleen" <email obscured>>
    Reply-To: <email obscured>" <email obscured>>
    Date: Friday, April 23, 2021 at 2:10 PM
    To: <email obscured>" <email obscured>>
    Subject: [EXTERNAL]Re: [ABRF Ab Forum] Shipping spleens

        Frances,
         I suggest you make a single cell suspension of the spleen put it in a
15ml screw cap tube and fill it with media + 10% FCS/FBS. Make sure there a
minimal air pocket and ship it on wet ice.
        I know if we put single cell suspension of spleens in media in an ice
bucket in the fridge overnight the cell viable does not change drastically.
One other thing, this is usually after rbc lysis.
        Kathy

        ________________________________
        From: Antibody Discussion Forum <email obscured>> on behalf of
Frances Weis-Garcia <email obscured>>
        Sent: Friday, April 23, 2021 1:12 PM
        To: <email obscured> <email obscured>>
        Subject: [ABRF Ab Forum] Shipping spleens

        Does anyone have experience shipping spleens?  Good or bad?

        A researcher has immunized mice and would like to ship me the spleen
for me to prep cells from and then fuse them.  We did a test run ... harvested
the spleen yesterday ... put it in a tube of media ... let it sit on ice packs
in a box overnight ... prepared a single cell suspension in parallel to a
freshly harvested spleen.  We recovered only 33% of the cells, relative to the
freshly prepared ones.  Lots of gooey tissue left behind after squeezing it
between 2 frosted slides but the cells were 74% viable (Fresh were 94%).

        I have never had good success with frozen splenocytes.

        Thoughts?
        ――
        View topic
https://nam04.safelinks.protection.outlook.com/?url=http%3A%2F%2Flist.abrf.org%2Fr%2Ftopic%2F1HS4gkcFyBdBuCuHARUfot&amp;data=04%7C01%7Ckbrundage%40hsc.wvu.edu%7C11a2225739cd4f04cd6308d9067b0b6d%7Ca2d1f95f851044248ae15c596bdbd578%7C0%7C0%7C637547947811266285%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C1000&amp;sdata=fPxDD1JVdiJOwld7j4rSACGBharCQcz3eaYk60uuhZQ%3D&amp;reserved=0
        Leave group <email obscured>?Subject=Unsubscribe


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EXTERNAL] Shipping spleens

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