I have been working on PC-3 and A549 cell culturing in DMEM+10%FBS for the last two weeks, and I have observed the slow growth of both cells. I need the expert's opinion on whether can I change the growth medium from DMEM to Ham's F-12K (ATCC recommended) during the culturing or the next passage phase, to improve the growth of these cells? I need your suggestions, Thanks in Advance!
Dear Dr. Faheem Askari
It is always better to use the culture media recommended by the vendor. If you have been working with PC-3 and A549 cells, I presume you are using the same media (DMEM) recommended by the vendor.
While most cell lines can grow in more than one culture medium, their characteristics are very much likely to alter when the medium is changed. So, using the same culture medium for cell growth in which the cells have been previously growing is advisable. It should not happen that in the process of changing the culture media you may lose the original characteristics of the cells.
Changing the media from DMEM to Ham's F-12K is not advisable. But if you want to improve the growth of these cells, keeping the FBS concentration constant at 10%, you may add extra nutrients in the form of supplements like
1. 2mM L-glutamine, is the major source of energy for cell proliferation.
2. Non-essential amino acids (NEAA) which the cells can synthesize via metabolic pathways such as glucoaminolysis, glycolysis, or the TCA cycle, the demand for these molecules sometimes becomes so great that efficient synthesis is difficult. So, adding supplemental non-essential amino acids to culture media will stimulate both growth and prolong the viability of the cells in culture. Usually, NEAA solution is provided as 100X concentration. You may add 1 mL NEAA solution to 99mL cell growth medium for culturing your cells.
3. 1mM sodium pyruvate helps improve the cell's ability to metabolize glucose in the media to produce energy. Sodium pyruvate solution may be added to cell culture media as an additional, easily accessible carbohydrate source in the cultivation of cells, especially for rapidly growing cells, such as tumor cells. Otherwise, it is generally not required for cells.
So, culturing cells in the same culture medium used by the vendor is recommended for best results.
Regards,
Malcolm Nobre
Dear Prof. Malcolm Nobre, Thank you so much for your valuable suggestions.
Actually, I took PC-3 cells from a lab at my institute and they told me that about 3 years before someone used and cultured these cells in DMEM media. Now, I tried to culture the same cells in DMEM media, but the cell's growth was not so good, and most of the cells remained detached from the flask surface. I found ATCC's recommendation to use Ham's F-12k media for the smooth growth of PC-3 cells, that's why I am doubtful to either change the media now to improve the cell growth or not. However, your suggestion to use extra nutrients is also meaningful. I will try this way to get my required results.
Dear Dr. Faheem Askari
I would suggest you first try using the supplements I mentioned. Hope the cells feel much happier in this environment and show you the results you expect.
Since the cells remain detached from the flask surface, I would consider the cells to be under environmental stress. What could cause environmental stress? Watch out for the following, if you haven't.
1. Supplementing media with extra nutrients, which you will be trying as mentioned above.
2. Contamination.
3. Inappropriate cell surface for attachment.
4. Improper gas mixture in the environment.
5. Temperature fluctuation in the incubator.
Check for those factors which you haven't.
Another possibility of cells undergoing stress would be the way you have thawed the cells from storage (either liquid nitrogen or -80 degree C). You have to follow a quick thawing process (less than 1 minute) in a 37 degree C water bath. So, in addition to the nutrients in the media, you need to check for other factors mentioned above that could be causing some sort of stress to the cells.
Best.
Dear Prof. Malcolm Nobre, Thank you so much for valuable suggestions. @Malcolm Nobre
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