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RPMI-1640 Medium Introduction

RPMI-1640 Medium is an artificial complex type medium. Moore and his colleagues originally developed this medium at Roswell Park Memorial Institute (RPMI) and based on the RPMI 1630 line of media that utilized a bicarbonate buffering system and alterations in the amounts of amino acids and vitamins. RPMI was found successfully for the cultivation of normal human and neoplastic leukocytes. It is now a popular general-purpose medium when properly supplemented and it is modified as follows with L-glutamine and phenol red and without  4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES).

Composition of RPMI-1640 Medium

RPMI 1640 Medium is different from other media since it contains the reducing agent glutathione and high concentrations of vitamins. This medium contains biotin, vitamin B12, and para-aminobenzoic acid (PABA), which are lacking in Eagle’s Minimal Essential Medium (EMM) or Dulbecco’s Modified Eagle Medium (DMEM). In addition, the vitamins inositol and choline are present in very high concentrations. RPMI 1640 Medium contains no proteins, lipids, or growth factors. Therefore, this medium requires supplementation, commonly with 10% Fetal Bovine Serum (FBS). It uses a sodium bicarbonate buffer system (2.0 g/L) and thus requires a 5–10% CO₂ environment to maintain physiological pH.

Modifications of RPMI-1640 Medium

RPMI-1640 Medium and its various modifications are RPMI-1640 Medium Dutch Modification, RPMI-1640 Medium HEPES Modification, RPMI-1640 Medium Modified, RPMI-1640 Medium Auto-Mod™ for Autoclaving, etc. RPMI-1640 Medium HEPES Modification is shown as an above image due to having Sigma code R5886 that indicates its identity.

 Composition of RPMI-1640 Medium HEPES Modification (R5886)

Inorganic Salts
Calcium Nitrate • 4H₂O
Magnesium Sulfate (anhydrous)
Potassium Chloride
Sodium Bicarbonate
Sodium Chloride
Sodium Phosphate Dibasic (anhydrous)
Amino Acids
L-Arginine
L-Asparagine (anhydrous)
L-Aspartic Acid
L-Cystine • 2HCl
L-Glutamic Acid
Glycine
L-Histidine
Hydroxy-L-Proline
L-Isoleucine
L-Leucine
L-Lysine • HCl
L-Methionine
L-Phenylalanine
L-Proline
L-Serine
L-Threonine
L-Tryptophan
L-Tyrosine • 2Na • 2H₂O
L-Valine
Vitamins
D-Biotin
Choline Chloride
Folic Acid
Myo-inositol
Niacinamide
p-Aminobenzoic Acid

D-Pantothenic Acid (hemicalcium)
Pyridoxine • HCl
Riboflavin
Thiamine • HCl
Vitamin B12
Other
D-Glucose
Glutathione (reduced)
HEPES
Phenol Red • Na
Add
L-Glutamine

Application of RPMI-1640 Medium

1. RPMI 1640 Medium has initially developed to culture human leukemic cells in suspension and as a monolayer. But it was also found suitable for a variety of mammalian cells, including HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
2. As a modification of McCoy′s 5A Medium, it was formulated to support lymphoblastoid cells in suspension culture, but it has since been shown to support a wide variety of cells that are anchorage-dependent.
3. Originally intended to be used with a serum supplement,  but RPMI 1640 was found to support several cell lines in the absence of serum. It has also been widely used in fusion protocols and in the growth of hybrid cells.
4. Manufacturers offer a variety of RPMI 1640 Medium modifications for a range of cell culture applications.

Keynotes

• In general, it’s always good to start  RPMI- 1640 for suspension cells and with MEM for adherent cells.
• Media should always be tested for sterility by placing it in a 37ºC CO₂ incubator for 3 days prior to utilization to ensure that the lot is contamination-free
• The medium should store at 4ºC.
• Several components of the medium are light-sensitive and therefore it should be stored in dark.
• RPMI-1640 plus 10 FBS % is useful for signaling studies in cell lines, Jurkat for lymphoblast morphological study of humans.
• RPMI-1640 plus 2mM glutamine and  10-20% FBS is useful for signaling studies in cell lines, HL60 for lymphoblast morphological study of humans.

References

1. https://www.thermofisher.com/order/catalog/product/11875135#/11875135
2. https://www.mpbio.com/rpmi-1640-with-l-glutamine-without-sodium-bicarbonate
3. https://www.sigmaaldrich.com/life-science/cell-culture/learning-center/media-formulations/rpmi-1640.html