Protocol Online logo
Top : New Forum Archives (2009-): : Tissue and Cell Culture

problems with adherent CHO culture - (May/04/2010 )

Hi
I have a problem with culturing adherent CHO cells, when I defrez a cell batch I can passage it for about a month without any problem, but after that cell growth decreases and they cannot attach to the flask any more and during a week they all are dead.
I never have this problem with other adherent cells, is there any help?
thanks in advance

-sara.r-

CHO cells are fairly easy cell line that grow rapidly and attach easily, but depending on the specific type of CHO cell line you are using they have different viable passage numbers. How old are the cells that you are bringing up? Are you consistently getting the same number of passages out of the cells before they stop adhering? There is a strong possibility the cells are starting to differentiate and they is why this keeps happening.

-sunshine1109-

I totally agree with Sunshine!

CHO cells are relatively hardy- however, once they get to a certain passage number, they are next to impossible to culture. I use CHO-k1 cells and those are great until passage 18-20-- then they start to get funky. Perhaps that would explain your observations.

-labrat612-

sunshine1109 on May 11 2010, 07:29 AM said:

CHO cells are fairly easy cell line that grow rapidly and attach easily, but depending on the specific type of CHO cell line you are using they have different viable passage numbers. How old are the cells that you are bringing up? Are you consistently getting the same number of passages out of the cells before they stop adhering? There is a strong possibility the cells are starting to differentiate and they is why this keeps happening.


thank you very much for your reply, acctually we bougth the cells from atcc, and I think they should send fresh cells, by the way what do you mean by differentiation of CHO cells? are immortalized cells capable of differentiation?

-sara.r-

ah ha! Cells from ATCC-- it is entirely possible that they sent you a bad lot. How long after did you start to notice these issues with the cells?

-labrat612-