clumping of MEFs in suspension

Jayakumar, R R.Jayakumar at RoswellPark.org
Fri Apr 8 10:39:35 EST 2005


MEF= Mouse embryo fibroblasts.  They don't clump. Check out for
mycoplasma contamination.   Some mutant cells show stress effects and
clump.  If they are clumped in huge clumps and floating, then they are
dead.  Throw them away. Check for viability with trypan blue exclusion
staining.
Best of luck



Jai

-----Original Message-----
From: owner-cellbiol at hgmp.mrc.ac.uk
[mailto:owner-cellbiol at hgmp.mrc.ac.uk] On Behalf Of Bob
Sent: Friday, April 08, 2005 1:24 AM
To: cellbiol at hgmp.mrc.ac.uk
Subject: Re: clumping of MEFs in suspension

On 7 Apr 2005 18:33:39 +0100, anette.huebner at umassmed.edu (Anette)
wrote:

>Hi everybody,
>I just finished a MEF 

What is an MEF?

Abbreviations and acronyms often have many meanings. Really helps to
give good info if you want good feedback.


>prep (one of many I have done so far), meaning I 
>just plated my cells 2 hours ago. The efficiency of the prep seemed 
>great.
>When I checked on them 10 min ago, I realized that the cells are 
>forming big clusters in suspensions. I have NEVER seen this before.
>Well, these are not WT MEFs, but a new mutant. I was wondering whether 
>this is something that happens for whatever reason, or, whether it 
>could be a phenotype.

Well, regardless of what MEF means, that is an interesting question.
You will find out by seeing if it is reproducible, with proper
controls. Perhaps a solution was not correct this time, or perhaps you
do have a novel phenotype of your mutations.

bob



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