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Media formulations for MDA-MB-231, BT-474 cells and Caco-2 cells - (Oct/12/2011 )

I am looking for some help with growing MDA-MB-231, Caco-2 and BT474 cells. I have obtained these cells recently from ATCC. I have a number of questions regarding media.

Currently I have BT474 cells growing in ATCC Hyrbicare 46-X media DMEM (4.5g/L glucose, L-glutamine, sodium pyruvate) with 10% FBS and 10mM sodium biocarbonate, as stated in the ATCC packet. I just thawed the BT474 cells and they have finally reached near confluencey, but some areas of the cells in the flask look granulated others do not. I can not find any good images of confluent BT474 cells online, could anyone help? Also this media I prepared with the powder seems to be slightly acidic (more orange) should this be adjusted, and how if so?

I am confused as to what media is best for the MDA-MB-231 cells. It seems ATCC reccomends L-15 media, but this is for incubation without CO2. I have seen a number of different formulations in the literature for growing these cells in CO2 incubators. Does anyone have a tried and true media formulation they are using for MDA-MB-231 grown in CO2 incubators?

For the Caco-2 cells, does anyone have a suggestion on media besides the ATCC formulation Eagle's Essential Medium? I am trying DMEM (4.5 g/L glucose, L-glutamine, sodium pyruvate), 10% FBS, 1% MEM-NEAA. I have seen several different formulations in the literature, again does anyone have a solid formulation they have worked with in the past? It seems that these cells maybe slightly more difficult to culture than the other two cell lines mentioned, is this true?

I am somewhat confused on the different types of DMEM out there. If the DMEM states it contains L-glutamine should more be added if the protocol calls for it? I know L-glutamine is labile, and thus can it hurt to add more over time to the media?

Should I use DMEM with lower glucose for any of these cell lines? Is it best to purchase base media and add your own supplements such as sodium pyruvate, L-glutamine or HEPES. This is where I am most unsure, in regards to the base media containing such components and then looking at a recipe which calls for the addition of such components.

Seems to be lots of information on media formulations but not so much on the actual nutrient requirements for each cell line...


For culture BT-474 cell line, u can try to add FBS up to 20%