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Problems with B-cell line BJAB


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#1 gobessi

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Posted 03 November 2004 - 04:16 AM

Hi!
I have some problems with a Burkitt's lymphoma cell line called BJAB....I'm maintaining the cells in standard normal conditions (RPMI, 10% FCS, L-glutamine, Pen/Strep antibiotics, Sodium Pyruvate), in flasks without shaking....
Some days ago I noted the formation of cell clumps that are very difficult to eliminate by pipetting or shaking (this clumps render tht counting of the cells very hard!!!...)....At the beginning of this phenomenon the cells grew well but I observed that the medium quickly became yellow.....and now, the cells do not grow as well as before.....
In parallel, I'm maintaining other B-cell lines that grow well, do not form these clumps, and the medium does not change the color as BJAB cells...
Is here someone that could explain me this phenomenon, and could help me to solve this problem?...Someone suggested me to add heparin....is it a good idea?

Thanks a lot!!!
Stefania

Edited by gobessi, 03 November 2004 - 04:29 AM.


#2 Jacen

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Posted 07 December 2004 - 08:16 AM

I think a yellow color in your cell media is indicative of a bacterial infection.
Is it cloudy as well?

#3 mez

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Posted 04 May 2010 - 07:56 AM

Hi!
I have worked with BJAB cells extensively and the problem is quite simple: the cells aggregate to form "giant" cells impossible to count.
The solution: grow your cells in RPMI + 10% FBS + 5-10 microM beta-mercaptoethanol. This will prevent formation of the giant cells. The cells will still clump but they can be broken up by pipetting up and down. Since clumping is favored by high density, do not grow past 1X10E6 cells/mL. Pass 1:10 every 3-4 days. Et voila! BJAB should be wonderful now!




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