JAWS II dendritic cells culture

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TheGrad
TheGrad's picture
JAWS II dendritic cells culture

Hi everyone,
I have been trying to culture the only mouse dendritic cell line available JAWS II, and was wondering if anyone had experience with that?
I am using the recommended ATCC medium:
alpha-mem with ribonucleosides, deoxyribonucleosides, 4mM Glutamine, 20% FBS 1mM sodium pyruvate and 5ng/ml GM-CSF.
The cells seem to be happy and healthy for about 3 weeks, proliferating at a slow but steady rate. And then they would all die at the same time (even separate plates) one day...
I haven't let them become more than ~60% confluency before splitting and been changing media every 2 days.

It's the third time it has happened to me, and I am not sure what's happening!

Any help/advice would be GREATLY appreciated!

Thank you!!

heehawmcduff
heehawmcduff's picture
Hi there,

Hi there,

I have never used this line but had a few general questions

First, have you had any problems with any other cell lines stored in the same incubator?

Second, is there a reason for you using 20% FBS (I am asking because this paper cultured them with 10% - http://iai.asm.org/cgi/reprint/IAI.01584-07v1)

Finally, when they die, has the media changed colour?

Best wishes

TheGrad
TheGrad's picture
Hey, Thanks for the quick

Hey, Thanks for the quick reply,

I am using 20% FBS because that is ATCC's recommended medium. But I've seen papers that use slightly different mediums, so I might try some of those.
My other cells in the same incubator/same conditions are fine.
And the media I culture JAWS cells in rarely ever changes color, not with confluency, or when they die...

Joanna

Oliwia
Oliwia's picture
Hi,

Hi,

Maybe after 3 weeks your media changed its properties? Maybe  L-glutamine or another unstable chemical broke down and it was extremely essential for this cell line? Try maybe to prepare new medium after this time or check if e.g. addition of L-glutamine or another suspicious reagent would make a difference.

Greetings,

PS. Please, let us know how the JAWSII are doing now? DO you know already what was the cause of their death? Good luck!

mikey mike
mikey mike's picture
I never had this problem.. I

I never had this problem.. I used exactly the same conditions as you...

I tried seeding these cells on 24 and 12 well plates.. and transfecting them... in some wells... the cells washes off very easily.... does anyone uses poly-l-lysine or poly-dl-lysine to coat your plates? which is better?

any advise would be greatly appreciated!