Fibroblasts are not good when culturing fused cells. They often will
dominate the culture and utilize media nutrients that your fused cells
need. Often presence of fibroblasts occurs when a substantially aged
animal was used as the splenocyte donor. To circumvent this, I've kept
the age of my immunized animals low prior to fusion.
-John
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Subject: Immuno Digest, Vol 15, Issue 8
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Today's Topics:
1. Re: adherent cells after fusion (Allison)
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Message: 1
Date: Fri, 13 Oct 2006 11:41:05 -0400
From: Allison <allison from nospam.com>
Subject: Re: [Immunology] adherent cells after fusion
To: immuno from net.bio.net
Message-ID: <BuKdne-W0_QPLrLYnZ2dnUVZ_t2dnZ2d from mcgill.ca>
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The adherent cells look spread out, as if they might be fibroblasts.
Not the kind of thing that can be dislodged by gentle pipetting (this I
already see with my Sp2/0 cells and I would consider it to be normal).
Allison
Haviland, David L wrote:
> Allison:
>> Funny you'd ask. I'm watching that very thing with my P3X based
fusions. Gentle pipetting or a smack on the side of the flask will
dislodge most. I guess they are just sticky after they have been
fused. However, I can't speak to the viability issue you bring up. The
one's in the media are alive and I increase the numbers by getting more
dislodged from the plastic.
>> David
>> -----Original Message-----
> From: immuno-bounces from oat.bio.indiana.edu on behalf of Allison
> Sent: Thu 10/12/2006 2:09 PM
> To: immuno from magpie.bio.indiana.edu> Subject: [Immunology] adherent cells after fusion
>> Any one have ideas on why sometimes I get adherent cells growing after
> doing a standard splenocyte - Sp2/0 fusion with PEG? This last time I
> had very few hybridoma clones, lots of dead/clumped cells, and afair
> number of adherent cells.
>> Thanks
>> Allison
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