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

MDA-MB-231 - stopped to grow. Help! - (Oct/05/2010 )

I need an advice. I've got new celllines MDA-MB-231/361 and they have grown in medium from lab which send this cells to me. But when I used our medium, a lot of cells died and they don't grow (or grow very slowly now). Should I wait? Can they got use to new medium (Probably new serum) or what should I do???
Please help, I am not much experienced.

-tadala-

If you are using a different medium to the lab that supplied the cells (e.g. they used RPMI1640 and you are using DMEM) you can wean them onto the new medium by switching from the other medium to your medium over several passsages, i.e. start out in 100% RPMI, at the next passage switch to 80% RPMI/20% DMEM, at the next passage, 60% RPMI/40% DMEM... etc untill you are totally in the medium of choice.

Be aware that some cell types, especially breast lines, don't like phenol red in the medium as it is an oestrogen mimic so you may have to use colourless medium for your experiments.

-bob1-

bob1 on Wed Oct 6 23:53:51 2010 said:


If you are using a different medium to the lab that supplied the cells (e.g. they used RPMI1640 and you are using DMEM) you can wean them onto the new medium by switching from the other medium to your medium over several passsages, i.e. start out in 100% RPMI, at the next passage switch to 80% RPMI/20% DMEM, at the next passage, 60% RPMI/40% DMEM... etc untill you are totally in the medium of choice.

Be aware that some cell types, especially breast lines, don't like phenol red in the medium as it is an oestrogen mimic so you may have to use colourless medium for your experiments.



I use the same medium (however from other company)and I suppose that it must have been serum which stressed cells. Now they look better. If one have to change serum (I mean take another batch), should try to mix two different serums (old and new) to avoid "schock"?

-tadala-

Ideally you would test each batch of serum to see if the cells will grow in a similar fashion (rate, morphogenesis, attachment etc.) in the new serum compared to the old one.

-bob1-

Did the cells arrive frozen? Sometimes cells recovering from cryopreservation take more than overnight to adhere to the bottom of a flask. Whenever I receive a new cell line, instead of discarding the medium after the first day of recovery, I transfer it to a new flask if a trypan blue stain on a media sample shows that some floating cells are still viable.

-sushik-

The cell are now O.K. I think it must have been shock after changed serum. However thay still need long time to attach to the culture flask.

anyway thank you for tips.

-tadala-