Hi Berrin

On a very basic level in our laboratory you would use DMEM for an adherent cell line and you would use RPMI for suspension cell lines. After that the additions such as alpha-MEM or high/low glucose is dependent on experience with the cells and what seems to work best for the scientist. For example some cell lines grow and flourish better in the presence of HEPEs. Also you need to think about your down the line uses for the cells. For example if you use your cell line for a fluorescence assay then Phenol red needs to be excluded from your culture medium as it has a high fluorescence and your background reading will be too high. All aspects are considered when deciding on the best media. I personally will research the cell line and see what is suggested and then try a few options to see what works the best for me.

Hi Berrin

On a very basic level in our laboratory you would use DMEM for an adherent cell line and you would use RPMI for suspension cell lines. After that the additions such as alpha-MEM or high/low glucose is dependent on experience with the cells and what seems to work best for the scientist. For example some cell lines grow and flourish better in the presence of HEPEs. Also you need to think about your down the line uses for the cells. For example if you use your cell line for a fluorescence assay then Phenol red needs to be excluded from your culture medium as it has a high fluorescence and your background reading will be too high. All aspects are considered when deciding on the best media. I personally will research the cell line and see what is suggested and then try a few options to see what works the best for me.

 Hi Angela,  I know some labs uses RPMI also for adherent cells.  But thank you for your help

Hi Bernin,

The best way to choose a proper medium for your cell culture is to go through the literature,( 5 0r 6 choices)and find the one that is most used.

Deborah Patinkin

Hi Berrin

Yes that is correct some do use RPMI for adherent cells, that's why I just say in my lab that's our basic rule of thumb. When ever you work with a new cell line you need to see what works best for you as Deborah stated use literature to see what the most used is and work form there.

Dear Deborah and Angela;

I allready do as you say. I check literature before I choose my Medium but It would be better if I do understand what is the real reason all those people use that kind of medium for that kind of cell type.

but anyway thank you very much. 

Hi Berrin

Please find the attached link. It has quite a good description on how to chose the culture media from a basic start to additives that can help and so forth.

http://www.labome.com/method/Cell-Culture-Media-A-Review.html

Thanks Angela . I was just printing the same thing. You are so kind and helpful. thank you very much

Dear Berrin,

There is not a universal rule about using a perticular media for a perticular cell type and even in literature people don't specify there standardized conditions and they hide some very crucial components thats why instead of trying the same culture media sometimes we don't get the expected results as promised by an article. Most of the cell lines are quiet flexible with any of the media you described above albeit if you will use a gud quality FBS (US origin) you will get more cell proliferation with good morphology and in this case dependency on choice of the media is minimal. You can try it practically in lab where one cell line you can grow in DMEM and after a passage or stock formation it can be adapted very well to RPMI or MEM. But in case of the primary culture, choice of the culture media is a very critical issue. eg. I am doing primary culture of stem cells and I tried almost 4-5 different types of media. Sometimes even there are variations in media constituents like MEM+L-glutamine+ Essential amino acids. In that case theoritically you should look on the type of the cells, the nutrient requirements, that cell type needs for growth in vivo and wheather those optimum combinations are available in company catalogues or not. Practically you should not only try different types of media in case of primary culture but in some cases u may also need to try different variations of a perticular media like DMEM+non essential amino acids+Glutamine or DMEM v/o L-glutamine but with non essential amino acids. It all depends on the type of the nutrition a perticular type of cell requires and the microenviorenment from which it has been isolated (not in case of cell lines as thet are immortalised). HAM'S F-12 AND OTHER NUTRIENTS are also required when your cells have specific requirements. So nutrient differe from cell to cell so as media changes depending on the suitability of the nutrients it can provide.. It can be then HGF for hepatocytes, RA for neural cells and so on.. That's why in case of specific cell types we need to shift to different media.  Hope it helps..!

I would like to thank all of you one by one.. Every answer was useful. Thank you very much

You choose the medium for the specific cell type that appears the most in the literature or in the articles resembling your experimental design.

just to share.. In most literature they use ALPHA-MEM for therapeutic use of hMSCs 

I think that it depend on your protocol and the kind of cells

Once you have established which basic medium and additional supplements you are going to use, I would recommend batch testing the serum you use. This is important for stem cells, since they are much more sensitive than cell lines. You can do this by requesting samples for batch testing from various different companies. They will send a small bottle of serum and reserve a stock of the same batch for you. You can then culture your hMSCs in the medium containing different sera for several weeks and observe cell morphology. Choose the best batch of serum, request that the company keeps the batch in stock for you for however long you need it (can be several years) and inform the other companies that you have chosen a different batch.

RPMI 1640 is also ok for Mesangial cells. I once culture this same cell line with RPMI 1640. If you prepare yourself, add 3g of Hepes/L and 2g of Sodium Carbonate. Generally, you may have to try different medium sometimes to choose the best for the optimum growth of the cells. You may also get the information from publication that have correlation with your line of research.

Hi Temitope Adeluci,

The concentrations you suggest, using 3g Hepes/L and 2g of sodium bicarbonate are for RPMI 1640 medium right? Do you have any information if these concentrations work in alpha MEM? I already use 2 g/L of sodium bicarbonate in the preparation of alpha MEM, but I would like to supplement my medium with HEPES, since when I put my cells at hypothermic temperatures (4 °C) the medium loses the optimal PH since it does not have the presence of CO2.

Thank you in advance

thank you very much for all your help. All of them was useful for me.

Please go through this link..

https://books.google.co.in/books?id=kiDpauK826MC&pg=PA98&lpg=PA98&dq=choosing+right+cell+culture+medium&source=bl&ots=7msFTsxCbG&sig=kaKSMTGF59NKLy-VzoMA9txzy4g&hl=en&sa=X&ved=0ahUKEwiMo-6frPXLAhWDCI4KHa1ADaIQ6AEIMzAB#v=onepage&q=choosing%20right%20cell%20culture%20medium&f=false

hello my friend

it is my exprience that using MEM alpha for mesenchymal stem cell sub culture is more effective than others.

Dear Abolfazl nosrati tirkani, can u please send catalogue of this? what supplements do u use?

Hi Dear Dr Mahmood.

May I Ask you to explane me which catalog or protocole do you need?

Hello

Attachments here will help you

Dear Berrin,

I just clarified my experience about mesenchymal SCs,

I used the alpha MEM because I recruited the MSCs for obtaining necessary cytokines,

Based on only my observations (beyond any scientific rules) The alpha MEM protects the MSCs from oxidative damage and keep them more uniform.

When choosing a cell culture medium, there are many important factors to be considered. On the underneath I made for you a short list for choosing appropriate cell culture medium:

• Appropriateness for species,
• research purpose,
• cell history and
• type

To know more details click here http://bit.ly/2wWIzHa

Human mesenchymal stem cell growth medium is also appropriate if you are working with human bone marrow MSCs. This can be obtained directly from dealer companies like Sigma. It is a complete medium.

It is important you make known the type of MSCs you intend to culture. These cells, though highly proliferative, but may require their type of media. I don't think you will need to use the same media for bone marrow MSC and Umbilical cord MSC?