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problem with COV318 ovarian cancer cell line


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#1 bongiwoman

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Posted 22 November 2013 - 11:48 AM

I am working with COV318 and noticed that the cells grow extremely slowly so I split them only 1:2 about once or twice a week. They are already passage 45 and they also seem to change their sizes and become bigger. P50 eg had 660000 cells in a confluent T75 flask and the freshly thawed P45 had 1.1 million and was only about 60% confluent I would say. Any clue whats going on...? Thanks so much!

#2 bob1

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Posted 23 November 2013 - 11:52 PM

Sounds like there is probably some sort of Hayflick limit approaching.  Don;t know about the cell line in particular though. 

 

COuld be some sort of hormone effect - are you using phenol red in your culture medium?  If so, it is an estrogen analogue, which means that ER+ cells will internalize the receptor and slow growth rates. 



#3 bongiwoman

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Posted 25 November 2013 - 02:56 AM

Hi Bob1, thanks a lot for your reply! I was also thinking that they might become sensescent - on Friday I seeded more cells than I counted today...and I had split them only 1:1.5.... A colleague told me also that cells can look like this after viral infection (by accident), but I dont know how to check for this. Obviously they are mycoplasma negative at least but I dont know how to work with those cells and how to figure out the problem. They grow in DMEM which indeed contains phenol red - do you think it could be worth trying with DMEM without phenol red? I also add NEAA to the medium but when I didn't add it they did not look any different.

#4 bob1

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Posted 25 November 2013 - 01:05 PM

I think it would be worth using phenol red free medium, though the original paper (pdf here) just used DMEM with l-glutamine (3 mM) and l-asparagine (1 mM).  I still think they are likely to be approaching the Hayflick limit, as they are essentially a primary line still, in which case there won't be anything you can do to prevent it happening.

 

Sigma sell them, so you could approach their technical support to see what they have to say.



#5 Nefeli

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Posted 22 January 2014 - 07:20 AM

Hi I am working with COV318 cells too. I just bought them from Sigma and they were Passage 44! 

bongiwoman what passage are your cells now if I may ask? Are they indeed senescent? I am kind of worried about the Hayflick limit myself since a lot of my cells are senescent since I revived them..




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