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What will happen if the cell culture medium is suddenly changed to another one?

DMEM MEM RPMI MCF-7

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#1 nemorat

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Posted 22 August 2012 - 07:24 AM

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.

#2 ascacioc

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Posted 22 August 2012 - 10:49 AM

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

#3 bob1

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Posted 22 August 2012 - 12:37 PM

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!

#4 albertross

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Posted 05 September 2013 - 11:00 AM

I know this post is a bit old, but what about using RPMI in some washes for glioblastoma cells cultured in "Complete" DMEM? I just joined a lab out of college, and was trained by a grad student who said the lab will try to save on expenses by using RPMI (apparently cheaper) for many of the washes that would otherwise by done with fresh DMEM. I know they're cancer cells and not as fickle as the hESCs I have some experience working with, but I was concerned that this could be a detrimental shock to the cells.



#5 albertross

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Posted 05 September 2013 - 11:07 AM

If it makes any difference, these aren't cells that have been passaged tons of times and are identical from lab to lab to lab. These are recovered from patients and the stocks kept to very low passage numbers, and we're trying to keep them as close to in vivo conditions as possible because of a collaborating lab's clinical trials.



#6 bob1

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Posted 05 September 2013 - 02:26 PM

If you want to go that far, why not wash in PBS? - much much cheaper.







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