FluxMPS™ RPMI 1640 Medium
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- See the table below
FluxMPS™ RPMI 1640 Medium
FluxMPS™ RPMI 1640 Medium is a 1X liquid, ready-to-use formulation of the classic medium developed in 1966 at the Roswell Park Memorial Institute by George E. Moore, Robert E. Gerner, and H. Addison Franklin, from which it derives its name. Originally formulated to support the growth of normal and neoplastic human leukocytes in suspension, RPMI 1640 remains the default basal medium for human lymphoid and hematopoietic cell culture. The 33-configuration family lets researchers select exactly which distinguishing components — L-Glutamine, Sodium Pyruvate, Sodium Bicarbonate, HEPES, and Phenol Red — are present in the finished medium, then filter every microfluidic-grade batch through the quadruple-stage FluxMPS™ system before it reaches an OoC, ToC, or LoC platform.
- 33 configurations: a standard bicarbonate-buffered RPMI 1640 line (17 configurations, including a special Folic Acid-free variant) and a HEPES-supplemented line (16 configurations), independently toggling L-Glutamine, Sodium Pyruvate, Sodium Bicarbonate, and Phenol Red
- Contains a high concentration of phosphate, amino acids, and vitamins essential for supporting cell growth and proliferation
- Bicarbonate buffering system requiring a 5-10% CO2 atmosphere to maintain a physiological pH, with an optional 25 mM HEPES-supplemented line for procedures outside a CO2 incubator
- 2.0 g/L standard glucose concentration
- Typical composition includes glucose, phenol red pH indicator, salts (NaCl, NaHCO3, Na2HPO4, KCl, MgSO4, Ca(NO3)2), amino acids (glutamine, arginine, asparagine, cystine, leucine, isoleucine, lysine, serine, aspartic acid, glutamic acid, hydroxyproline, proline, threonine, tyrosine, valine, histidine, methionine, phenylalanine, glycine, tryptophan, reduced glutathione), and vitamins (inositol, choline chloride, para-aminobenzoic acid, folic acid, nicotinamide, pyridoxine HCl, thiamine HCl, calcium pantothenate, biotin, riboflavin, cyanocobalamin)
- Purified through FluxMPS™ quadruple-stage 0.1 micron / 0.04 micron filtration, engineered for organ-on-chip (OoC), tissue-on-chip (ToC), and lab-on-chip (LoC) microfluidic platforms
- Versatile medium supporting many adherent and suspension cell types, including lymphocytes, Jurkat cells, HeLa cells, bone marrow cells, hybridomas, and carcinomas
- Available in 500 mL and 1000 mL sizes; store at 2-8°C protected from light; every standard concentration, buffer, and additive is customizable on request
- Concentration1X
- Glucose2.0 g/L
- BufferingSodium bicarbonate/CO2, optional 25 mM HEPES
- Configurable supplementsGlutamine / Pyruvate / Bicarbonate / HEPES / Phenol Red
- Special variantFolic Acid-free (DCP-RPMI-F1X)
- FiltrationFluxMPS™ quadruple-stage, 0.04 micron final
- Sizes500 mL, 1000 mL
- Storage2-8°C, protected from light
At-a-Glance Supplement Matrix
Check the supplement(s) your protocol requires, press Search, and every matching configuration lights up below. Click the catalog number or the View button to go straight to that product page.
| Name | Cat No. | L-Glutamine | Sodium Pyruvate | Sodium Bicarbonate | HEPES | Phenol Red | Special | Product Page |
|---|---|---|---|---|---|---|---|---|
| RPMI 1640 | DCP-RPMI1X | check | check | check | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine | DCP-RPMI-Q1X | remove | check | check | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o Sodium Pyruvate | DCP-RPMI-P1X | check | remove | check | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o Sodium Bicarbonate | DCP-RPMI-B1X | check | check | remove | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o Phenol Red | DCP-RPMI-R1X | check | check | check | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Sodium Pyruvate | DCP-RPMI-QP1X | remove | remove | check | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Sodium Bicarbonate | DCP-RPMI-QB1X | remove | check | remove | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Phenol Red | DCP-RPMI-QR1X | remove | check | check | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o Folic Acid | DCP-RPMI-F1X | check | check | check | remove | check | No Folic Acid | Viewarrow_forward |
| RPMI 1640, w/o Sodium Pyruvate, Sodium Bicarbonate | DCP-RPMI-PB1X | check | remove | remove | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o Sodium Pyruvate, Phenol Red | DCP-RPMI-PR1X | check | remove | check | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o Sodium Bicarbonate, Phenol Red | DCP-RPMI-BR1X | check | check | remove | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Sodium Pyruvate, Sodium Bicarbonate | DCP-RPMI-QPB1X | remove | remove | remove | remove | check | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Sodium Pyruvate, Phenol Red | DCP-RPMI-QPR1X | remove | remove | check | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Sodium Bicarbonate, Phenol Red | DCP-RPMI-QBR1X | remove | check | remove | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o Sodium Pyruvate, Sodium Bicarbonate, Phenol Red | DCP-RPMI-PBR1X | check | remove | remove | remove | remove | Viewarrow_forward | |
| RPMI 1640, w/o L-Glutamine, Sodium Pyruvate, Sodium Bicarbonate, Phenol Red | DCP-RPMI-QPBR1X | remove | remove | remove | remove | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES | DCP-RPMIH1X | check | check | check | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine | DCP-RPMIH-Q1X | remove | check | check | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Sodium Pyruvate | DCP-RPMIH-P1X | check | remove | check | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Sodium Bicarbonate | DCP-RPMIH-B1X | check | check | remove | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Phenol Red | DCP-RPMIH-R1X | check | check | check | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Sodium Pyruvate | DCP-RPMIH-QP1X | remove | remove | check | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Sodium Bicarbonate | DCP-RPMIH-QB1X | remove | check | remove | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Phenol Red | DCP-RPMIH-QR1X | remove | check | check | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Sodium Pyruvate, Sodium Bicarbonate | DCP-RPMIH-PB1X | check | remove | remove | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Sodium Pyruvate, Phenol Red | DCP-RPMIH-PR1X | check | remove | check | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Sodium Bicarbonate, Phenol Red | DCP-RPMIH-BR1X | check | check | remove | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Sodium Pyruvate, Sodium Bicarbonate | DCP-RPMIH-QPB1X | remove | remove | remove | check | check | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Sodium Pyruvate, Phenol Red | DCP-RPMIH-QPR1X | remove | remove | check | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Sodium Bicarbonate, Phenol Red | DCP-RPMIH-QBR1X | remove | check | remove | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o Sodium Pyruvate, Sodium Bicarbonate, Phenol Red | DCP-RPMIH-PBR1X | check | remove | remove | check | remove | Viewarrow_forward | |
| RPMI 1640 + HEPES, w/o L-Glutamine, Sodium Pyruvate, Sodium Bicarbonate, Phenol Red | DCP-RPMIH-QPBR1X | remove | remove | remove | check | remove | Viewarrow_forward |
RPMI 1640 (Roswell Park Memorial Institute Medium 1640)
RPMI 1640 is a basal synthetic cell culture medium developed in 1966 at the Roswell Park Comprehensive Cancer Center (then Roswell Park Memorial Institute), Buffalo, New York, by George E. Moore, Robert E. Gerner, and H. Addison Franklin. It was originally formulated to support the growth of normal and neoplastic human leukocytes in suspension, and it remains the default basal medium for all human lymphoid and hematopoietic cell culture more than six decades after its introduction. RPMI 1640 is structurally a modification of the RPMI 1630 series of media — itself a derivative of McCoy's 5A — and carries several compositional features absent from Eagle-lineage media (BME, MEM, DMEM): the highest inositol content of any classical medium (35 mg/L), calcium nitrate in place of calcium chloride as the sole calcium source, the highest phosphate concentration of any classical medium (800 mg/L Na2HPO4), reduced glutathione, vitamin B12, biotin, and para-aminobenzoic acid (PABA). These features reflect its design for non-adherent blood cells rather than substrate-dependent fibroblasts and epithelial lines.
Origins and Development
Roswell Park Memorial Institute and the Suspension Culture Problem
During the early 1960s, the growth of human lymphoid cells in vitro remained a significant technical challenge. Existing media — Eagle's BME, MEM, DMEM, and their derivatives — had been optimized for adherent cell types grown in monolayer and did not sustain long-term proliferation of primary human lymphocytes or lymphoblastoid cell lines in suspension. The underlying problem was nutritional: lymphoid cells in suspension experience a different metabolic microenvironment from adherent fibroblasts, with higher demands for inositol (a phospholipid membrane precursor), specific vitamins (B12, biotin), and antioxidant protection (glutathione), and they are more sensitive to calcium-driven adhesion to plastic surfaces. Moore and colleagues at Roswell Park Memorial Institute addressed these limitations by systematically modifying the RPMI 1630 medium series, an in-house set of formulations already in use at the institute, to improve support for lymphoid suspension cultures. The resulting medium, designated 1640 in the institute's sequential numbering system, incorporated substantially elevated inositol, a full complement of water-soluble vitamins including biotin, B12, and PABA, reduced glutathione as an antioxidant, calcium nitrate as the calcium source (reducing surface-adhesive calcium availability), and a dibasic-phosphate-dominant buffering salt set with high total phosphate.
The 1967 Publication
Moore, Gerner, and Franklin published the foundational paper as "Culture of normal human leukocytes," JAMA, 199(8): 519-524, February 20, 1967. The paper described the successful long-term culture of normal peripheral human leukocytes in RPMI 1640 supplemented with serum, providing a practical tool for lymphocyte biology and establishing the medium's suitability for immunological research. In the Citation Classics retrospective, Moore noted that the medium number 1640 was simply the sequential identifier of the final formulation from an extended screening series of modifications, analogous to Eagle's BME-to-MEM development path and the CMRL group's Medium 199 and CMRL 1066 series. The medium was developed one year before the publication, in 1966, and the 1967 paper reports the result of that development work.
Relationship to Eagle-Lineage Media
RPMI 1640 sits at the end of an indirect lineage from Basal Medium Eagle, distinct from the direct BME-to-MEM-to-DMEM path. It shares the Eagle-derived nutritional philosophy but diverges substantially in its calcium source, phosphate level, vitamin complement, and antioxidant provision — reflecting its design target of non-adherent lymphoid suspension culture rather than adherent fibroblasts and epithelial lines.
RPMI 1640 Lineage: McCoy's 5A to RPMI 1630 to RPMI 1640
RPMI 1640 sits at the end of an indirect lineage from Basal Medium Eagle. The developmental chain is:
- BME (Eagle, 1955) — Eagle lineage baseline; the original defined medium of 13 amino acids, 9 vitamins, 6 ionic species, and glucose.
- McCoy's 5A Medium (McCoy, Lui, & Brander, 1959) — a BME-derived modification developed at the University of Texas for the culture of Novikoff hepatoma cells; contains non-essential amino acids, nucleosides, and additional components beyond the Eagle essential set.
- RPMI 1630 series (Moore et al., early 1960s) — in-house modifications of McCoy's 5A at Roswell Park; the direct parent of RPMI 1640.
- RPMI 1640 (Moore, Gerner & Franklin, 1966/1967) — the final optimized formulation with elevated inositol, calcium nitrate, high phosphate, glutathione, and a full vitamin B12/biotin/PABA complement, described in the 1967 JAMA paper as the successor to the RPMI 1630 series and designated 1640 as the sequential identifier of the final formulation from an extended screening series.
RPMI 1640 Standard Formulation Reference
The tables below reproduce the canonical RPMI 1640 formulation as published in the source formulation record. All values are as stated in the source; no additional numeric values are implied for any specific catalog number beyond what is stated here and in the supplement matrix above.
| Ingredient | mg/L |
|---|---|
| Calcium nitrate tetrahydrate [Ca(NO3)2.4H2O] | 100.000 |
| Magnesium sulfate, anhydrous (MgSO4) | 48.840 |
| Potassium chloride (KCl) | 400.000 |
| Sodium bicarbonate (NaHCO3) | 2000.000 |
| Sodium chloride (NaCl) | 6000.000 |
| Sodium phosphate dibasic, anhydrous (Na2HPO4) | 800.000 |
| Amino Acid | mg/L |
|---|---|
| Glycine | 10.000 |
| L-Arginine (free base) | 200.000 |
| L-Asparagine (free base) | 50.000 |
| L-Aspartic acid | 20.000 |
| L-Cystine 2HCl | 65.000 |
| L-Glutamic acid | 20.000 |
| L-Glutamine | 300.000 |
| L-Histidine (free base) | 15.000 |
| L-Hydroxyproline | 20.000 |
| L-Isoleucine | 50.000 |
| L-Leucine | 50.000 |
| L-Lysine hydrochloride | 40.000 |
| L-Methionine | 15.000 |
| L-Phenylalanine | 15.000 |
| L-Proline | 20.000 |
| L-Serine | 30.000 |
| L-Threonine | 20.000 |
| L-Tryptophan | 5.000 |
| L-Tyrosine disodium salt dihydrate | 29.000 |
| L-Valine | 20.000 |
| Vitamin | mg/L |
|---|---|
| D-Biotin | 0.200 |
| Choline chloride | 3.000 |
| D-Calcium pantothenate | 0.250 |
| Folic acid | 1.000 |
| myo-Inositol | 35.000 |
| Niacinamide (Nicotinamide) | 1.000 |
| para-Aminobenzoic acid (PABA) | 1.000 |
| Pyridoxine hydrochloride | 1.000 |
| Riboflavin | 0.200 |
| Thiamine hydrochloride | 1.000 |
| Vitamin B12 (Cyanocobalamin) | 0.005 |
| Other Components | Amount |
|---|---|
| D-Glucose (Dextrose) | 2000.000 mg/L (~11.1 mM) |
| Glutathione (reduced, GSH) | 1.000 mg/L |
| Sodium bicarbonate (NaHCO3) | 2000.000 mg/L (2.0 g/L) |
| Phenol red sodium salt | 5.000 mg/L |
Sodium pyruvate is absent from the standard RPMI 1640 base, though many commercial pre-formulated RPMI variants add sodium pyruvate at 1 mM (110 mg/L) as an additional energy source. RPMI 1640 contains no exogenous lipid components and no nucleosides or nucleotide precursors in the standard base formulation. No ferric nitrate; iron is supplied entirely through serum. Calcium nitrate (rather than calcium chloride) provides calcium at approximately 0.42 mM while contributing nitrate ions rather than chloride, deliberately chosen to minimize calcium-dependent cell-substrate adhesion in support of non-adherent suspension culture of lymphoid cells. Sodium phosphate dibasic at 800 mg/L (~5.6 mM) is the highest phosphate concentration of any classical medium. NaCl was reduced to 6000 mg/L, compensated osmotically by the elevated phosphate and bicarbonate, maintaining an overall osmolality in the 255-310 mOsm/kg range appropriate for human blood cell culture.
RPMI 1640 vs. Classical Media
| Feature | RPMI 1640 | Standard MEM | High-Glucose DMEM | IMDM | Medium 199 |
|---|---|---|---|---|---|
| Developer & year | Moore, Gerner & Franklin - Roswell Park, 1966/1967 | Eagle - NIH, 1959 | Dulbecco & Freeman, 1959 | Iscove & Melchers, 1978 | Morgan, Morton & Parker - CMRL, 1950 |
| Lineage | McCoy's 5A → RPMI 1630 → RPMI 1640 | BME → MEM | BME → DMEM | DMEM derivative | CMRL (original) |
| Amino acids (entries) | 20 (19 of 20 standard, no Ala; + Hyp) | 13 (essential only) | 15 (+ Gly, Ser) | 17 | 21 (19 proteinogenic, no Asn; + cystine, + Hyp) |
| Vitamins (count) | 11 | 8 | 8 | 10 | 17 |
| Vitamin B12 | 0.005 mg/L | Absent | Absent | 0.013 mg/L | Absent |
| Biotin | 0.200 mg/L | Absent | Absent | Present | 0.010 mg/L |
| PABA | 1.000 mg/L | Absent | Absent | Absent | 0.050 mg/L |
| myo-Inositol | 35.000 mg/L | 2.000 mg/L | 7.200 mg/L | 7.200 mg/L | 0.050 mg/L |
| Vitamin B6 form | Pyridoxine HCl | Pyridoxal HCl | Pyridoxine HCl | Pyridoxal HCl | Both pyridoxal + pyridoxine HCl |
| Reduced glutathione | 1.000 mg/L | Absent | Absent | Absent | 0.050 mg/L |
| Glucose | 2000 mg/L (~11.1 mM) | 1000 mg/L (~5.5 mM) | 4500 mg/L (25 mM) | 4500 mg/L (25 mM) | 1000 mg/L (~5.5 mM) |
| Sodium pyruvate | None (standard); optional add | None | Optional (110 mg/L) | 110 mg/L | None |
| Calcium source | Ca(NO3)2.4H2O 100 mg/L | CaCl2 anhydrous 200 mg/L | CaCl2.2H2O 265 mg/L | CaCl2.2H2O 265 mg/L | CaCl2.2H2O 265 mg/L |
| Calcium concentration | ~0.42 mM | ~1.8 mM | ~1.8 mM | ~1.8 mM | ~1.8 mM |
| Iron source | None (serum-dependent) | None | Fe(NO3)3.9H2O 0.1 mg/L | KNO3 (transferrin-dependent) | Fe(NO3)3.9H2O 0.72 mg/L |
| Phosphate | Na2HPO4 800 mg/L (~5.6 mM) | NaH2PO4 140 mg/L (~1.0 mM) | NaH2PO4.H2O 125 mg/L (~0.9 mM) | NaH2PO4 113 mg/L | NaH2PO4 122 mg/L |
| NaHCO3 | 2000 mg/L (2.0 g/L) | 2200 mg/L | 3700 mg/L | 3024 mg/L | 2200 mg/L |
| CO2 required | 5-10% | 5-10% | 5-10% | 5-10% | 5-10% |
| pH (with NaHCO3) | 7.0-7.4 | 7.3-7.9 | ~7.2 | ~7.2 | 7.2 |
| Osmolality | 255-310 mOsm/kg | 290-330 mOsm/kg | 320-355 mOsm/kg | ~290-320 mOsm/kg | 310-350 mOsm/kg |
| Serum supplement | 10% FBS (5-20%) | 10% FBS | 10% FBS | 10% FBS | 10% FBS |
| Primary application | Human lymphocytes, PBMCs, Jurkat, hybridomas, P. falciparum | Fibroblasts, neurons, WI-38, MRC-5 | HEK293, MEFs, high-density adherent | Lymphocytes, hematopoietic cells, serum reduction | Chick fibroblasts, oocyte IVM, virology |
| Culture format | Suspension dominant, monolayer capable | Monolayer | Monolayer | Suspension and monolayer | Monolayer and organ culture |
RPMI 1640: Moore, Gerner & Franklin, Roswell Park, 1966/1967. MEM: Eagle 1959. DMEM: Dulbecco & Freeman 1959. IMDM: Iscove & Melchers 1978. Medium 199: Morgan, Morton & Parker, CMRL, 1950.
Why FluxMPS™
Quadruple-Stage Purity
Every FluxMPS™ RPMI 1640 configuration is purified to 0.04 microns — finer than any ready-to-use cell culture media currently available — before it ever reaches your chip.
Engineered for Microfluidics
Designed from the ground up for Organ-on-Chip (OoC), Tissue-on-Chip (ToC), and Lab-on-Chip (LoC) platforms, where the medium itself is part of the instrument.
Particulate & Aggregate Removal
The quadruple-stage architecture eliminates microscopic particulates and protein aggregates that silently block micro-channels and disrupt laminar flow — a particular concern for suspension lymphoid cultures.
Optical Clarity
Optical clarity supports real-time imaging and integrated biosensing on Organ-on-Chip platforms without background interference.
Regulatory-Aligned Foundation
Formulated to support FDA-recognized physiological modeling standards, providing a validated, reproducible media foundation as downstream data moves toward regulatory scrutiny.
Translational Research Ready
A single, consistent media platform supporting drug discovery, toxicology screening, and translational research from bench through preclinical modeling.
Quadruple-Stage Filtration
Every FluxMPS™ RPMI 1640 batch passes through a four-stage filtration train before final fill, engineered specifically for microfluidic cell culture applications.
-
1
0.1 µmPre-filtration Stage One
Bulk particulate reduction prior to sterile filtration.
-
2
0.1 µmPre-filtration Stage Two
Second-pass pre-filtration to protect the downstream sterile filters and extend their working life.
-
3
0.04 µmSterile Filtration Stage One
First sterile-filtration pass at 0.04 microns — finer than any ready-to-use cell culture media currently available.
-
4
0.04 µmSterile Filtration Stage Two
Second sterile-filtration pass, eliminating microscopic particulates and protein aggregates before final fill.
Built for Continuous Flow
Zero-clogging performance across complex micro-channel geometries and long-term automated perfusion studies running continuously for weeks.
Validated Cell Lines & Applications
RPMI 1640 is a versatile medium that supports the cultivation of many adherent and suspension cell types, such as lymphocytes, Jurkat cells, HeLa cells, bone marrow cells, hybridomas, and carcinomas. It is often used in a wide range of biological research applications, such as cell biology studies, virus propagation, and toxicity testing, and is typically supplemented with fetal bovine serum or serum replacements to provide additional growth factors and nutrients necessary for optimal cell growth.
Human Leukocytes & PBMCs
Primary human peripheral blood mononuclear cells (PBMCs), isolated by density gradient centrifugation and cultured in RPMI 1640 + 10% FBS, with stimulation by PHA, PMA/ionomycin, or anti-CD3/anti-CD28 in standard 72-hour T-cell activation assays. Fresh human lymphocytes were the original Moore et al. (1967) application; human monocytes and bone marrow / hematopoietic progenitor suspension cultures are also routinely maintained in RPMI 1640.
Jurkat, Raji, Ramos, HL-60, K562, THP-1, U937, Daudi
RPMI 1640 is the default maintenance medium for virtually all human lymphoid, leukemic, and lymphoblastoid cell lines, including Jurkat (T-cell leukemia), Raji (Burkitt's lymphoma; ATCC standard validation line), HL-60 (promyelocytic leukemia), K562 (myelogenous leukemia), THP-1 (monocytic leukemia), U937 (histiocytic lymphoma), Daudi (Burkitt's lymphoma), and the myeloma lines RPMI 8226 and U266, all in suspension culture with 10-20% FBS.
Hybridoma Culture & Monoclonal Antibodies
RPMI 1640 is the standard medium for murine and human hybridoma culture and cell fusion protocols: SP2/0 and NS0 myeloma fusion partners and derived hybridomas are maintained in RPMI 1640 + 10-20% FBS, with HAT/HT-supplemented RPMI 1640 used for post-fusion selection and hybridoma supernatant monitored via the phenol red color shift as cell density rises.
Plasmodium falciparum In Vitro Culture
RPMI 1640 enabled the first continuous in vitro cultivation of Plasmodium falciparum (Trager & Jensen, 1976), using human erythrocytes maintained in RPMI 1640 with human serum under a low-oxygen, high-CO2 atmosphere. The medium's elevated phosphate and inositol, lower calcium, and glutathione content were compatible with human erythrocyte physiology, and the system remains the global standard for P. falciparum culture, with Albumax I now used as the serum replacement in most laboratories.
Carcinoma Lines, Giardia & Nematode Biology
Many carcinoma lines, including HeLa, can be grown in RPMI 1640 in addition to their primary Eagle-lineage media. RPMI 1640 supplemented with L-cysteine supports Giardia lamblia trophozoite maintenance and murine spleen cell viability in co-culture immune assays, and a published modification supports Litomosoides carinii filarial nematodes.
General Suspension & Research Applications
RPMI 1640 supports various cell types including HeLa, BHK-21, 293, HEP-2, HT-1080, MCF-7, fibroblasts, and primary rat astrocytes, and is used in a wide range of biological research applications such as cell biology studies, virus propagation, and toxicity testing; any mammalian cell grown in suspension or semi-suspension can be evaluated in RPMI 1640 as a standard starting point.
RPMI 1640 in Physiologic Media Research & Practical Considerations
Physiologic Media Comparator
RPMI 1640 serves as one of the two primary "conventional supraphysiologic comparator" media (alongside high-glucose DMEM) in the physiologic media literature. The Cantor (2019) Trends in Cell Biology review identifies RPMI 1640 alongside DMEM as exhibiting amino acid concentrations that markedly differ from human plasma, and a glucose level (11.1 mM) that, while lower than high-glucose DMEM, remains supraphysiologic relative to blood (~5 mM). RPMI 1640 is the standard reference medium against which plasma-like formulations (HPLM, Plasmax) are benchmarked in metabolic cancer research and tumor metabolism studies.
Commercial Variants & Modifications
RPMI 1640 + HEPES (25 mM) is the dominant formulation for Plasmodium falciparum culture, PBMC assays requiring extended bench time, and T-cell activation assays. High-glucose (4.5 g/L) variants serve hybridoma production with increased energy demand; the ATCC-modified formulation (30-2001) adds L-glutamine, HEPES, sodium pyruvate, elevated glucose, and reduced NaHCO3; and an Advanced RPMI 1640 variant adds ethanolamine, ascorbic acid, glutathione, insulin, transferrin, lipid-rich albumin, and trace elements to reduce serum requirement by 50-90%.
Practical Considerations
L-Glutamine at 300 mg/L (2.05 mM) degrades spontaneously to pyroglutamate and ammonia during liquid-phase storage; liquid RPMI 1640 is frequently supplied without L-glutamine, and stable dipeptide alternatives (L-alanyl-L-glutamine) are available. Phenol red at 5 mg/L is half the concentration used in DMEM and MEM, reducing background absorbance in plate reader assays; phenol-red-free RPMI 1640 is available for estrogen-receptor and luminescence-based cytotoxicity assays.
Cold Storage Caution & Serum Reduction
Research has documented that RPMI 1640 is particularly deleterious during cold storage of adherent kidney cell lines (e.g., Vero-B4), an injury attributable to the combination of low calcium (~0.42 mM) and high phosphate (~5.6 mM); this limitation is specific to hypothermic storage and does not affect standard culture conditions, but RPMI 1640 should not be used as an organ preservation medium for renal cell preparations. The base formulation's glutathione, biotin, B12, and PABA content reduces the serum requirement compared to MEM and DMEM for many lymphoid lines, with some lymphoblastoid lines maintained at 2-5% FBS in RPMI 1640 versus 10% FBS required in DMEM.
Frequently Asked Questions
Verified References
- Moore, G.E., Gerner, R.E. & Franklin, H.A. (1967). Culture of normal human leukocytes. JAMA, 199(8): 519-524. PMID 4960081. DOI 10.1001/jama.1967.03120080061009.
- Trager, W. & Jensen, J.B. (1976). Human malaria parasites in continuous culture. Science, 193(4254): 673-675. PMID 781840.
- Cantor, J.R. (2019). The Rise of Physiologic Media. Trends in Cell Biology, 29(11): 854-861. PMC7001851. PMID 31623927.
- Yao, T. & Asayama, Y. (2017). Animal-cell culture media: History, characteristics, and current issues. Reprod. Med. Biol., 16(2): 99-117. PMC5661806.
- Moore, G.E., Gerner, R.E. & Franklin, H.A. (1968). New York State Journal of Medicine, 68: 2054. Follow-up characterization of RPMI-series media.
- Moore, G.E., et al. (1976). TCA Manual, 3: 503.
FluxMPS™ — Precision Cell Culture Media for Microphysiological Systems
Built for the architecture of the future. Not the flask of the past.
Traditional cell culture media were formulated for static well plates and flasks — environments that tolerate impurities, precipitates, and particle loads that would immediately compromise a microfluidic system. FluxMPS™ was designed from the ground up for Organ-on-Chip (OoC), Tissue-on-Chip (ToC), and Lab-on-Chip (LoC) platforms, where the medium itself is part of the instrument.
Purity That Protects Your Platform
FluxMPS™ is purified to 0.04 microns — finer than any ready-to-use cell culture media currently available. At this level, the microscopic particulates and protein aggregates that silently block micro-channels, disrupt laminar flow, and generate false biological signals are eliminated before the media ever reaches your chip.
The result: your platform stays operational, your data stays clean, and your biology drives the result — not your media.
Engineered for Flow, Not Just Growth
The name FluxMPS™ reflects its core design principle. Every component is optimized for consistent, laminar flow performance across:
- Complex micro-channel geometries
- Capillary-bed and vascular simulations
- Long-term automated perfusion studies running continuously for weeks
Zero-clogging performance is not a feature — it is the baseline specification.
Applications & Performance
| Application | What FluxMPS™ Delivers |
|---|---|
| Microfluidics | Stable shear stress; no channel blockage |
| Metabolic Tracing | Ultra-pure matrix with no contaminant interference |
| Long-term Perfusion | Consistent formulation stability over weeks of continuous flow |
| Organ-on-Chip | Optical clarity for real-time imaging and integrated biosensing |
Regulatory Foundation
FluxMPS™ is formulated to support FDA-recognized physiological modeling standards, providing a validated, reproducible media foundation for drug discovery, toxicology screening, and translational research. When your downstream data needs to stand up to regulatory scrutiny, your upstream media cannot be an afterthought.
The Bottom Line
Microfluidic platforms are precision instruments. They require precision inputs.
FluxMPS™ is the only ready-to-use cell culture medium engineered specifically to meet that standard — protecting your chip, your cells, and your science.
