culturing Jurkat T-cells - (Nov/30/2007 )

for some reasons we plan to start with Jurkat T-cells but have no experience; I need some basic information:

adherent or not?
are there sublines, and which is commonly used?
generation time?
difficulty of handling?
other special information?

thanks for any help

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Non-adherent, there are sublines that I know of but those are transfected ones.
Generation time is around 24 hours if I'm not mistaken and they are easy to handle.

What do you need to do with them that you can't do with other cells?

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thanks vairus; we are interested in PKCtheta which is known to be expressed in T-cells; PKCtheta is raely expressed in most other types of cells;

still another question: are Jurkat cells oncogenic or just non-oncogenic immortal?

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Dear "The Bearer",

Our group have been growing Jurkats for many years. They are very easy to maintain if conditions are kept constant i.e.

Use the best quality FBS/FCS and BATCH TEST IT.
Start the cells in Tissue Culture flasks, for example a T75.
Never "over split" the cells i.e. 1:10- 1:20 split ratio. BUT AT FIRST when they are recovering from cryopreservation, go for 1:2/1:4 splits.
Once they are established, grow the cells in Techne Stirrer Bottles.....this allow you to grow 100's of millions of cells for your experiments.

Our groups reference papers:

PNAS Beltan et al Dec. 19 2000, Vol 97, No.26, pp 14602-14607

PNAS Beltran et al June 25 2002, Vol 99, No.13, pp 8892-8897

Hope this is useful.

Rhombus

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thank you Rhombus, very helpful information; I will start as you advice

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Are these cells different than the Jurkat cells? Just curious because Jurkat cells too have the same properties described as above.

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Hi,

I am growing Jurkat cells for the first time too and although I think things are going well, I just wanted to check on a few little things:

1) The RPMI media that I am using appears to be slightly orange now that I have refrigerated it in 4º, not the pink color it was when I received it. I know that this is an indication of a PH change, is this normal and okay?

2) I am growing the cells in 75cm2 vented flasks and, aside from the reccomended volume limitations on this particular flask, is there an optimum volume that is best for maintaining these cells?

3) I am maintaining them at a density between 1 and 2 x 10(6), this is more dense that is reccomended but they seem to be doing okay, is there any problem with this?

4) When I am preparing to mix the FBS with the media, can I put the FBS in a 37º water bath to defrost? Also, I am adding Pen/Strep antibiotic-can I defrost this in 37º waterbath?

Thanks in advance.

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Dear Elephantman,

In our experience it is best to keep the cells at between 250,000 and 750,000 cells/ml. WE DO NOT USE ANTIBIOTICS.... it masks POOR TECHNIQUE. And a s stated previously, the cells are best grown in TECHNE stirrer bottles...this keeps them in suspension and more importantly STOPS THEM AGGREGATING.

If you look after them properly they are one of the easiest cells to grow in large numbers.

Kindest regards

Rhombus

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