I culture THP-1 cells in RPMI (10% FBS, 1% Pyruvate, 1% non essential amino acids and 1% Pen/Strep). I have been differentiating the THP's into neural macrophages, however I have had 2 batches of cells that have been cultured without pyruvate and amino acids. The media has been changed but the cells are less reactive and instead of 'sitting down', they are clumping.
I have differentiated the cells in both RPMI and ADMEM and they behave the same in both. The above is the only obvious difference but I have been advised it shouldn't make a difference.
Any comments or advice is very welcome.
Andrea
Hello Andrea Daniels-Morgan
THP-1 is a very sensitive cell line. Culturing THP-1 cells without pyruvate and non-essential amino acids could make a difference in cells’ growth and behavior. Their response to external stimuli could be altered.
Sodium pyruvate is used by cells as an easily accessible carbohydrate source. Additionally, it is involved with amino acid metabolism and initiates the Kreb's cycle.
The use of non-essential amino acids is basically to maintain cell growth. Most cells can synthesize non-essential amino acids (NEAA) through glucoaminolysis, glycolysis, or the TCA cycle. When concentrations of these amino acids are low, cells need to consume more glucose and glutamine in the media, and produce excess by-products, which might affect cell growth. Adding NEAA into cell culture media can mitigate the potential side effects caused by self-production of these non-essential amino acids.
I would recommend the addition of pyruvate and non-essential amino acids in the cell culture media (RPMI 1640 + 10% FBS) for culturing THP-1 cells.
Best.
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