chicken lymphoma cell line

[Aawara Chowdhury]

In <mailman.604.1175549162.5139.methods from net.bio.net>,
 Rebecca Pickin <RPickin from cvm.msstate.edu> wrote:

> 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.)  

You haven't mentioned the name of the cell-line.  I have grown both chicken 
DT40 and DT95 cells.  These are bursal lymphoma cell-lines that work well 
for homologous recombination.

The conditions that I've used are considerably different that those
you've listed below:

> 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.

Media:
DMEM/HG with 4 mM L-glutamine
10% tryptose-phosphate broth
10% fetal bovine serum
5% chicken serum

(no antibiotics, no fungizone/amphotericin B, no Hepes)

5% CO2 incubator, 37oC.  Incidentally, I used similar conditions to
grow other avian cell-lines, such as the quail QT6 and QT35 fibroblasts,
and line-0 CEFs. 

In terms of growing them, I always maintained them in "log", so that
I would split them when they reached a density between 3x10e5 cells/ml,
to 5x10e5 cells/ml.  I would seed new dishes at 8x10e4 cells/ml, and
would split them every 3 - 4 days.  The media was replaced every time
I split the cells.

Good luck!
AC
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