3 replies to this topic

[MariaS]

Hello, we are trying to culture SKBR-3 cells. We've been using McCoy 5A (from Gibco, Invitrogen) +10% FBS for a while now. We used to split them in 3 twice a week, with one additional feeding in between. Recently (~ since 2 months ago), we notice that the cells stop growing and die in approximately 2 weeks after they are thawed unless they are fed every day. And even then they stop growing and do not adhere to the bottom of the flask. We did not change anything in the way we culture them. Has anybody experienced anything like that with this line or with this company's media? Thanks!

[bob1]

Have you recently got a new batch of FBS in?  This sort of thing is quite common with new batches of serum, especially if it hasn't been tested on the cell line you are using...

[MariaS]

No, we still have the same batch of FBS, and the same batch of antibiotic. The only thing changed is the lot of the media.

[bob1]

Are you using phenol-red free medium?  If not, you may be seeing the effects of long term selection for cells with an active estrogen receptor.