2 replies to this topic

[nemorat]

Hello, everyone.

Does any know what will happen to the cells, or what damage will do to the cells when cell culture media is suddenly changed to another one, like DMEM to RPMI?

We are using MCF-7 cells, and the old lab protocol is to culture it in RPMI, which is not the recommended culture medium for MCF-7 by ATCC.  Suddenly we got a new batch that has been maintained in MEM, and we found the ones in either MEM or DMEM have the same morphology, but the ones that have been maintained in RPMI have different morphology.

Then I got a flask of MCF-7 cells, but I didn't know they are in DMEM. So I followed the old protocol and changed the cell culture medium to RPMI unknowingly. The morphology became different one day after changing to RPMI.

I have some questions.

1. Does it matter if MCF-7 has been maintained in MEM, DMEM, or RPMI? I have read some posts on the website, but the answer is still not clear to me.

2. What happened to those MCF-7 cells whose culture medium was suddenly changed from DMEM to RPMI? Can we still use them?

3, I froze some cells whose culture medium was suddenly changed from DMEM to RPMI. Can I thaw them into DMEM or MEM? Will the morphology change back? Or is it that I cannot use them at all?

Thank you.

[ascacioc]

The different media have different components that can up-regulate/down-regulate certein proteins -->differentiation. However, MCF-7 are terminally differentiated cancer cells (as far as I remember they are the breast cancer cells) so I wouldn't see why they wouldn't be the same in between changes of the media. So my answer is: you can change media of cancer cells and nothing will happen. But use the ATCC recommendations: there were some people who tested several media and decided that a certain media is better. I always take their recommendations and my life was easier because of them:)

Andreea

[bob1]

Sudden changes also induce stress on the cells as they have to get used to different levels of certain compounds in the medium.  This means that some of the cells will die, and you will be left with a population of cells that are resistant to those changes - you just changed the cell line, so that it is no longer what you thought it was...

To change cells into different media, you need to do this slowly over several passages using increasing amounts of the new medium and decreasing amounts of the old medium.  This is referred to as "weaning" the cells into a new medium.  After this process, you should characterize the cells for morphology, growth rate, expression of various genes to ensure that the cells have not changed significantly from the parent line.  This is also important to ensure that your experiments are not going to be affected by the changes!