RPMI 1640 Medium is unique from other media because it contains the reducing agent glutathione and high concentrations of vitamins. RPMI 1640 Medium contains biotin, vitamin B12, and PABA, which are not found in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. In addition, the vitamins inositol and choline are present in very high concentrations. This RPMI formulation has been modified by ATCC to contain high glucose, low sodium bicarbonate, sodium pyruvate, HEPES, and low L-glutamine. RPMI 1640 Medium contains no proteins, lipids, or growth factors. Therefore, RPMI 1640 Medium requires supplementation, commonly with 10% Fetal Bovine Serum (FBS). RPMI 1640 Medium uses a sodium bicarbonate buffer system (1.5 g/L) and therefore requires a 57% CO2 environment to maintain physiological pH.

cGMP manufacturing and quality system：
RPMI 1640 Medium is manufactured at a cGMP-compliant facility located in Grand Island, New York. The facility is registered with the FDA as a medical device manufacturer and is certified to ISO 13485 standards.