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| Product Name | RPMI 1640 Medium, with L-Glutamine, without Sodium Bicarbonate, Liquid, Sterile |
| Catalog No. | ATR-CCR-0039 |
| Description | Sterile liquid RPMI 1640 (Roswell Park Memorial Institute) basal medium formulated with L-glutamine (2 mM) but without sodium bicarbonate, for use in 5% CO2 atmosphere after bicarbonate supplementation or for CO2-independent applications. RPMI 1640 is the most widely used medium for human lymphoid cells, hybridomas, leukemia and lymphoma cell lines, and many adherent cancer cell lines. Originally developed for culture of human normal and neoplastic leukocytes, RPMI 1640 supports the growth of suspension cultures including Jurkat, HL-60, THP-1, U937, K562, Raji, Daudi, and many hybridoma cell lines, as well as adherent lines including HeLa, MCF-7 (with insulin), and HCT116. The medium uses a bicarbonate/CO2 buffer system supplemented with sodium phosphate for additional buffering. This formulation without pre-added sodium bicarbonate allows users to add the precise amount needed for their specific culture conditions or to use the medium with alternative buffer systems (HEPES) for CO2-independent applications. |
| Intended Use | Basal culture medium for: human and animal lymphoid and myeloid suspension cell lines (Jurkat, THP-1, HL-60, U937, Raji, Daudi, K562); human and mouse hybridomas for monoclonal antibody production; leukemia and lymphoma research; human tumor cell lines (HeLa, MCF-7, HCT116, A549, PC-3); primary human lymphocytes (with IL-2 supplementation); and as the base medium for formulation of complete growth medium with serum and supplements. |
| Principle / Technology | RPMI 1640 contains 20 amino acids (including reduced glutathione and para-aminobenzoic acid), 11 vitamins (biotin, choline, folate, nicotinamide, pantothenate, pyridoxine, riboflavin, thiamine, vitamin B12, i-inositol, and para-aminobenzoic acid), 6 inorganic salts, glucose (2 g/L, 11.1 mM), and phenol red pH indicator. L-glutamine (2 mM) is pre-added for immediate use. Sodium bicarbonate is not included, allowing custom addition (typically 2.0 g/L for 5% CO2). The phosphate buffer system provides some pH buffering capacity independent of CO2, and reduced glutathione serves as an antioxidant. |
| Detection Method | 1) Warm medium to 37 C before use; 2) Add sodium bicarbonate (typically 2.0 g/L for 5% CO2) as needed; 3) If using for CO2-independent culture, add HEPES (10-25 mM final) and adjust pH to 7.2-7.4 with NaOH; 4) Supplement with fetal bovine serum (typically 5-10% v/v for most cell lines) and optional antibiotics (penicillin-streptomycin); 5) Sterile-filter through 0.22 um filter if adding non-sterile supplements; 6) Use complete medium within 2-4 weeks when stored at 2-8 C protected from light. |
| Sample Type | Ready-to-use sterile liquid basal medium; requires supplementation with serum and optional antibiotics before use with cells; pre-warm to 37 C for cell culture. |
| Performance Range / Specifications | pH: 6.8-7.2 (without NaHCO3, equilibrated with air); after NaHCO3 addition and CO2 equilibration: pH 7.2-7.4; osmolality: 250-290 mOsm/kg (without serum); glucose: 2.0 g/L (11.1 mM); L-glutamine: 2.0 mM; endotoxin: <0.25 EU/mL; sterility: negative (no growth 14-day test, USP <71>); mycoplasma: negative. |
| Sensitivity / LOD | Not applicable — basal cell culture medium. |
| Specificity | Formulated specifically for lymphoid and myeloid cell culture; can support many adherent cell types but DMEM is often preferred for adherent fibroblast and endothelial cells. RPMI 1640 has lower calcium (0.42 mM vs. 1.8 mM in DMEM) and magnesium concentrations, which may affect some adherent cell attachment. |
| Reaction Conditions / Protocol | Cell culture at 37 C in 5% CO2 humidified atmosphere; medium pH color indicator: phenol red — orange-red at pH 7.4, yellow (acidic) when medium is spent, magenta (alkaline) when CO2 is low. |
| Components / Formulation | RPMI 1640 Medium (1x liquid), without sodium bicarbonate, with L-glutamine (2 mM), phenol red, sterile-filtered (0.1 um), 500 mL bottle. |
| Storage Conditions | Store at 2-8 C protected from light (light degrades L-glutamine, vitamins, and phenol red); do not freeze. |
| Shelf Life | 12 months from manufacture at 2-8 C unopened; after opening and supplementation: 4-6 weeks at 2-8 C when handled aseptically. |
| Package Specifications | 500 mL, 1,000 mL sterile PETG or glass bottles. |
| Product Form | Clear, red-orange sterile liquid (phenol red indicator); no particulates. |
| Quality Control | Each lot: pH 6.8-7.2 (without NaHCO3); osmolality 250-290 mOsm/kg; glucose 1.8-2.2 g/L; L-glutamine 1.8-2.2 mM; endotoxin <0.25 EU/mL; sterility: negative (bacterial and fungal, 14 days); mycoplasma: negative (PCR and culture); cell growth test: Jurkat and THP-1 cells achieve >=20-fold increase in viable cell density over 72 h with 10% FBS. |
| Key Features | RPMI 1640 without sodium bicarbonate (customizable); L-glutamine pre-added; sterile-filtered; endotoxin tested; phenol red pH indicator; supports lymphoid and myeloid suspension cells; widely used standard formulation. |
| Purity | All components: cell culture grade or USP grade; water: WFI quality; endotoxin <0.25 EU/mL; trace elements as specified in original Moore formulation. |
| Concentration | 1x liquid medium; glucose 2.0 g/L; L-glutamine 2 mM; amino acids and vitamins per RPMI 1640 standard formulation (Moore et al., 1967). |
| Activity / Unit Definition | Cell growth promotion: verified with Jurkat (>=20-fold in 72 h, 10% FBS). |
| Molecular Weight | Glucose: 180.16 g/mol; L-glutamine: 146.14 g/mol. |
| Source / Origin | All components: synthetic or fermentation-derived; water: WFI quality (distillation or reverse osmosis + electrodeionization); no animal-derived components in basal medium; manufactured in ISO-certified facility under cGMP for medical devices. |
| pH Range / Optimal pH | pH 6.8-7.2 without NaHCO3; after NaHCO3 and 5% CO2: pH 7.2-7.4; pH decreases as cells metabolize (lactic acid accumulation) — medium change indicated when phenol red turns yellow. |
| Shipping Conditions | Ambient temperature (medium stable for 2 weeks at RT); cold pack for extended transit recommended. |
| Expiration Date / Stability | 12 months at 2-8 C; after opening, 4-6 weeks at 2-8 C; L-glutamine degrades at 37 C and in light (half-life approximately 3-4 weeks at 4 C, 1 week at 37 C) — for long-term culture, supplement with fresh L-glutamine (2 mM) after 2-3 weeks of storage. |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic use. Not for human or animal injection. Manufactured under ISO 13485. Not GMP-certified for clinical cell manufacturing. |
| Compatibility | Compatible with standard cell culture incubators (37 C, 5% CO2, >95% humidity). Compatible with fetal bovine serum (FBS), calf serum, horse serum at 5-20%. Compatible with antibiotics: penicillin-streptomycin, gentamicin, amphotericin B. Compatible with selection antibiotics: G418 (Geneticin), hygromycin B, puromycin, zeocin. CO2-independent use: supplement with 10-25 mM HEPES and adjust pH to 7.2-7.4 with NaOH; cultures can be maintained in closed flasks without CO2 atmosphere. For adherent cells that require calcium for attachment (many epithelial lines), RPMI 1640 contains 0.42 mM Ca(NO3)2 — lower than DMEM (1.8 mM CaCl2); if poor attachment is observed, consider DMEM or supplement RPMI with additional CaCl2. |
| Recommended Buffer System | Bicarbonate/CO2 buffer system (bicarbonate added by user, typically 23.8 mM / 2.0 g/L) with phosphate buffer supplement; phenol red pH indicator (10 mg/L). |
| Application Notes / Precautions | Before use, add 2.0 g/L sodium bicarbonate (23.8 mM) for standard 5% CO2 culture. Adjust sodium bicarbonate concentration for different CO2 levels: 1.5 g/L for 3% CO2, 2.2 g/L for 7.5% CO2, 2.7 g/L for 10% CO2. The medium bottle should be warmed to 37 C before opening to minimize pH shift from CO2 absorption. Supplement with L-glutamine if medium has been stored for >3 weeks at 4 C or if heated to 37 C for extended periods. For hybridoma culture: RPMI 1640 supplemented with 10-20% FBS, 1 mM sodium pyruvate, and 50 uM 2-mercaptoethanol supports optimal growth. RPMI 1640 is susceptible to light degradation; protect bottles from light during storage and incubation (wrap in aluminum foil if needed). |
| Batch-to-Batch Consistency | pH 6.8-7.2; osmolality 250-290 mOsm/kg; glucose 1.8-2.2 g/L; glutamine 1.8-2.2 mM; cell growth verification for every lot. |
For research use only, not for clinical use.
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