Hello everyone!
I have trouble with a cell line, namely the human kidney carcinoma Caki-2 cell line. Another lab sent us a few criovials of cells, with their growing conditions, but I can't grow them at all. They are incredibly slow, most of them die as soon as thawed, and their morphology is not the one that I see on photos on the net.
The lab that sent me the criovials grow them in DMEM high glucose with 10%FBS, plus Pen-strep and L-Glutamine. They told me that Caki-2 should be a pretty easy cell line that does not need much effort, but I can't grow them at all. All my other cell lines grow ok in my incubator, so I think that that's not the problem. Does anyone have any experience with this cell line and can help me? Maybe some small detail that I am missing?
Thank you!
Hi,
I used to work with them years ago, but I can't remember any problems with it. There are several things you can check:
- how long does it take you, after defrosting, to start worrying? Usually, frosting is a stressful process and even if done quickly [defrosting], it takes a while for cells to recover.
- how is with sodium pyruvate in the medium?
- was shipment done correctly?
- is there anything suspicious in the medium? Pen-strep isn't the most effective mix. Do you add any anti-mycotic?
I am aware you might check them already, however, I hope I could help.
Good day!
Maybe you look at this protocol, there use RPMI-1640 medium
Hello and thanks for the replies!
Wojciech Witarski it has been two weeks now and they are still really slow, and when I defrosted them, the vast majority was already dead. Shipment, from what I know, was done correctly. Cells arrived in dry ice and were still frozen. I stored them in liquid nitrogen until actual use. Sodiu pyruvate is present. Maybe they do not need it? Nothing suspicious in the medium, it's not a contamination problem.
I now split them once every week. From what I can understand, you should be able to split them at least 2-3 times every week.
Alexandr Chernov we decided to keep some of them in RPMI, but they are still incredibly slow, although visually slightly better.
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