chicken lymphoma cell line

Jayakumar, R via methods%40net.bio.net (by R.Jayakumar from roswellpark.org)
Mon Apr 2 17:11:57 EST 2007


HI.. Becky
I am aware that some cell lines prefer the higher 41C (have you tried
lower temperatures though I very much doubt that this is the problem).
But these are also extremely sensitive to CO2 conditions too at that
temperature.  Please enlighten me as to how you determine the CO2
concentrations in your incubator before I go any further.  
Jai
 

-----Original Message-----
From: methods-bounces from oat.bio.indiana.edu
[mailto:methods-bounces from oat.bio.indiana.edu] On Behalf Of Rebecca Pickin
Sent: Monday, April 02, 2007 4:22 PM
To: methods from magpie.bio.indiana.edu
Subject: chicken lymphoma cell line

Hello all!

I am currently trying to grow chicken lymphoma-like suspension cell line
and having no luck at all.  (I have done google searches for this
particular line and have come up with nothing.)  The lab is working with
the individual who created these cells, but still having no luck.  I
have tried my cell culture tricks and have run out of ideas.  I was just
wondering if any of you had any tricks to growing challenging cells in
culture.  

These have been recommended to grow in a humidified chamber at 41oC and
5% CO2.  They prefer to be between 200,000 cells/ml and 800,000 cells/ml
and are in a medium which contains the following:
RPMI, 20% FBS, 1x PNS (pen/strep), 1:500 amphotericin B, 25mM HEPES, 4mM
L-glutamine, 1mM Sodium Pyruvate.  We were checking them and feeding
them every 3 days.

Currently the cells are all dead and we are trying to get more.  Does
anyone know if there might be a better medium formulation or a
substitution for an ingredient which might be better than what I've
listed?  Any magic tricks?  I've prayed to the lab gods and did my
special "please-let-this-work" dance to no avail.  

Any and all help would be appreciated.

Thanks in advance!
Becky







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