RPMI Medium 1640, IC-2032-S
Product Introduction
RPMI Medium 1640 is a classic and widely used basal cell culture medium originally developed by Moore and colleagues at Roswell Park Memorial Institute (RPMI) for the long-term culture of human lymphoid and suspension cells. It has since become one of the most commonly employed media for a broad range of mammalian cell types including T and B lymphocytes, hematopoietic cells, hybridomas, and numerous human cancer cell lines. RPMI 1640 is characterized by its unique formulation containing a relatively high phosphate concentration, reducing agents such as reduced glutathione, and a comprehensive panel of vitamins including inositol at elevated levels. The medium is buffered with a sodium bicarbonate/CO₂ system, supplemented with essential and non-essential amino acids, and formulated without HEPES to maintain physiological pH under standard 5% CO₂ incubation conditions. This product is supplied as a sterile liquid medium requiring only supplementation with fetal bovine serum and antibiotics according to specific cell line requirements. Its broad applicability across suspension and adherent cell culture systems has established RPMI 1640 as a foundational medium in immunology, oncology, and cell biology research.
Product Features
1. Classic RPMI 1640 Formulation: Trusted and extensively cited basal medium formulation optimized for lymphoid, hematopoietic, and suspension cell culture.
2. Comprehensive Amino Acid Profile: Contains all essential and non-essential amino acids including L-glutamine (300 mg/L) and L-cystine, supporting robust cell growth and metabolism.
3. Enriched Vitamin Supplementation: Includes elevated inositol (35 mg/L), choline, and a full panel of B-complex vitamins essential for cellular enzymatic reactions and proliferation.
4. Reducing Agent Included: Formulated with reduced glutathione (1 mg/L), providing antioxidant protection for sensitive suspension and lymphoid cell types.
5. Versatile Cell Type Compatibility: Suitable for a wide range of suspension and adherent cell types including human peripheral blood lymphocytes, hybridomas, myeloma, and leukemia cell lines.
Specifications
Size: Available in 500 mL and 20¡Á500 mL
Formulation: See detailed formulation table below
pH: 7.2-7.4
Osmolality: 260-290 mOsm/kg
Formulation Table (mg/L)
Inorganic Salts:
Ca(NO₃)₂¡¤4H₂O: 100
MgSO₄ (anhydrous): 48.84
KCl: 400
NaHCO₃: 2000
NaCl: 6000
NaH₂PO₄ (anhydrous): 800
Amino Acids:
L-Alanyl-L-Glutamine: -
Glycine: 10
L-Arginine HCl: 241.86
L-Asparagine¡¤H₂O: 56.82
L-Aspartic Acid: 20
L-Cystine¡¤2HCl: 65.2
L-Glutamic Acid: 20
L-Glutamine: 300
L-Histidine HCl¡¤H₂O: 20.22
L-Hydroxyproline: 20
L-Isoleucine: 50
L-Leucine: 50
L-Lysine HCl: 40
L-Methionine: 15
L-Phenylalanine: 15
L-Proline: 20
L-Serine: 30
L-Threonine: 20
L-Tryptophan: 5
L-Tyrosine¡¤2Na¡¤H₂O: 28.83
L-Valine: 20
Vitamins:
Biotin: 0.2
Choline Chloride: 3
D-Calcium Pantothenate: 0.25
Folic Acid: 1
Niacinamide: 1
Para-Aminobenzoic Acid: 1
Pyridoxine HCl: 1
Riboflavin: 0.2
Thiamine HCl: 1
Vitamin B12: 0.005
myo-Inositol: 35
Other Components:
D-Glucose: 2000
Glutathione (reduced): 1
HEPES: -
Phenol Red: 5
Penicillin G Sodium: -
Streptomycin Sulfate: -
Storage and Stability
Storage Conditions: Store at 2¡æ to 8¡æ, protected from light.
Shelf Life: The product is stable for 18 months from the date of manufacture when stored unopened as directed. Once opened, use within 4-6 weeks when stored properly and handled aseptically.
Protocol (For Reference Only)
Important: Use aseptic technique when handling medium to prevent contamination. Warm medium to 37¡æ before use for optimal cell growth.
1. Preparation of Complete Medium: Supplement RPMI Medium 1640 with fetal bovine serum to a final concentration of 5-10% and add antibiotics (e.g., penicillin-streptomycin) as required for specific cell lines. Mix gently by swirling.
2. Medium Pre-warming: Place the required volume of complete medium in a 37¡æ water bath or incubator for 15-30 minutes before use to minimize thermal shock to cells.
3. Suspension Cell Culture: Seed suspension cells (e.g., Jurkat, HL-60, hybridomas) at an appropriate density in complete RPMI 1640 and incubate at 37¡æ in a humidified 5% CO₂ incubator. Subculture when cells reach appropriate density.
4. Adherent Cell Culture: For adherent cells, seed cells in tissue culture-treated flasks or plates and incubate at 37¡æ with 5% CO₂. Aspirate spent medium and replace with fresh pre-warmed complete RPMI 1640 every 2-3 days.
5. Monitoring: Observe phenol red color indicator daily to assess pH changes; medium should appear pink-red at physiological pH and turn yellow-orange as acidity increases from cellular metabolism.
Precautions
1. Store medium at 2-8¡æ and protect from light to prevent degradation of light-sensitive components including L-glutamine and phenol red.
2. Avoid repeated warming and cooling cycles; aliquot medium into smaller volumes for daily use to maintain sterility and component stability.
3. Supplemented complete medium should be used within 2-4 weeks when stored at 4¡æ as glutamine degrades and antibiotic activity diminishes over time.
4. This formulation does not contain HEPES; ensure adequate CO₂ atmosphere (5-10%) for proper pH maintenance during culture.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What cell types are typically cultured in RPMI 1640?
A1: RPMI 1640 is the preferred medium for human lymphoid cells, hematopoietic cells, hybridomas, and many leukemia and lymphoma cell lines. Common cell lines cultured in RPMI 1640 include Jurkat, HL-60, THP-1, Raji, and U937. It is also widely used for peripheral blood mononuclear cell (PBMC) culture.
Q2: Does this medium contain L-glutamine?
A2: Yes. This formulation contains L-glutamine at 300 mg/L. For long-term culture, additional L-glutamine supplementation may be beneficial as glutamine degrades over time at 4¡æ and 37¡æ. Consider stable glutamine substitutes for extended culture periods.
Q3: Why is there no HEPES in this formulation?
A3: RPMI 1640 is traditionally formulated without HEPES, relying on the sodium bicarbonate/CO₂ buffer system for pH regulation. This is optimal for standard 5% CO₂ incubator conditions. If extended handling outside the incubator is required, HEPES may be supplemented separately.
Q4: What is the significance of reduced glutathione in RPMI 1640?
A4: Reduced glutathione serves as an antioxidant, protecting cells from oxidative stress. This is particularly important for lymphoid and suspension cell types, which may be more sensitive to oxidative damage compared to adherent fibroblast-like cells.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of culture conditions including serum concentration and supplementation is recommended for specific cell lines.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data resulting from product use or storage.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-2032-S
Product Name: RPMI Medium 1640
Size: 500 mL / 20¡Á500 mL
Price:
500 mL: CNY ¥200.00 / USD $20.00 / EUR €24.00 / JPY ¥3600.00
20¡Á500 mL: CNY ¥4000.00 / USD $400.00 / EUR €480.00 / JPY ¥72000.00
2023 Version
