[Immunology] RE: Immuno Digest, Vol 15, Issue 8

Hardaway, John Crawford via immuno%40net.bio.net (by hardawayj from missouri.edu)
Sat Oct 14 12:11:01 EST 2006


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)


----------------------------------------------------------------------

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>
Content-Type: text/plain; charset=us-ascii; format=flowed

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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> 
> 


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