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Qustion about MDA-MB-157 cells culture - (Jun/10/2009 )

I am carrying MDA-MB-157 cells (freshly from ATCC) and found they growed very very slowly. I just followed some groups to use the medium of DMEM+10%FBS and culture them in 5% CO2 + 95% air incubator.
Anyone knows what medium can be used and help the cells grow faster? Appreciate for any useful information. Thank you.

-Jerry-

ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 100%
Temperature: 37.0�C


As you got them from the ATCC, they will have been grown in the above medium... to use DMEM you need to wean them on to it slowly over several passages by using different mixtures of the two media.

-bob1-

It makes sense. Thank you very much, Bob1.



bob1 on Jun 11 2009, 09:07 PM said:

ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Atmosphere: air, 100%
Temperature: 37.0�C

As you got them from the ATCC, they will have been grown in the above medium... to use DMEM you need to wean them on to it slowly over several passages by using different mixtures of the two media.

-Jerry-

Hi, 

Bob1 can you tell me what kind mixture of the DMEM/10%FBS and L-15/10%FBS to use to get them to grow on 100% DMEM/10%FBS?

I was thinking DMEM:L-15 ratios as follows:  Pn 1.5:8.5, Pn+1 3:7, Pn+2 4.5:5.5, Pn+3 6:4, Pn+4 7.5:2.5, Pn+5 9:1, Pn+6 100% DMEM. Do you think this is a good or should I do 10% increments over more passages? Since the cells have to be re-fed 1x before passing the cells (splitting)  is it safe to increase the% of  DMEM between passages so that they are on 100% DMEM sooner? Or, should i just increase the % of DMEM only after splitting? 

 

Thanks a bunch in advance!
 

-AtxTom-

It is fine to do it with medium changes as well as during splitting, it is mostly so that the cells have time to adjust to the different components and concentrations in the different media.  Those ratios are fine, but a slower process will be less likely to give changes in the cells.

 

After you have done the weaning process, you should test to ensure that the cells still behave like they did before the weaning - growth rates, morphology, expression of genes of interest etc.

-bob1-