RPMI-1640

With L-Glutamine
Without Glucose and Sodium bicarbonate

Product Code: AT150

Product Description :
Roswell Park Memorial Institute (RPMI) media are a L-Leucine 50.000
series of media developed by Moore et al for the culture L-Lysine hydrochloride 40.000
of human normal and neoplastic cells in vitro. RPMI L-Methionine 15.000
1640 is the most commonly used medium in the series. L-Phenylalanine 15.000
A modification of McCoy's 5A medium, the medium was L-Proline 20.000
specifically designed to support the growth of human L-Serine 30.000
lymphoblastoid cells in suspension culture. Presently L-Threonine 20.000
the medium is extensively used for a wide range of L-Tryptophan 5.000
anchorage dependant cell lines. The medium needs to L-Tyrosine sodium salt 28.830
be supplemented with 5-20% fetal bovine serum. The L-Valine 20.000
medium is also known to support growth of cells in the VITAMINS
absence of serum. Choline chloride 3.000
D-Biotin 0.200
AT150 is RPMI 1640 with L-glutamine. It does not D-Ca-Pantothenate 0.250
contain glucose. Users are advised to review the literature Folic acid 1.000
for recommendations regarding medium supplementation Niacinamide 1.000
and physiological growth requirements specific for Pyridoxine hydrochloride 1.000
different cell lines. Riboflavin 0.200
Thiamine hydrochloride 1.000
Composition : Vitamin B12 0.005
i-Inositol 35.000
Ingredients mg/L p-Amino benzoic acid (PABA) 1.000
INORGANIC SALTS
Calcium nitrate tetrahydrate 100.000
Directions :
Glutathione reduced 1.000
Magnesium sulphate anhydrous 48.840 1. Suspend 8.4gms in 900 ml tissue culture grade water
Phenol red sodium salt 5.300 with constant, gentle stirring until the powder is completely
Potassium chloride 400.000 dissolved. Do not heat the water.
Sodium chloride 6000.000 2. It may be necessary to lower the pH to 4.0 with 1N HCl
Sodium phosphate dibasic anhydrous 800.000 to completely dissolve this product. After it has dissolved
AMINO ACIDS completely, the pH can be raised to 7.2 with 1N NaOH prior
Glycine 10.000 to the addition of sodium bicarbonate.
L-Arginine hydrochloride 241.000 3. Add 2.0gms of sodium bicarbonate powder (TC230) or
L-Asparagine 50.000 26.7ml of 7.5% Sodium bicarbonate solution (TCL013) for
L-Aspartic acid 20.000 1 litre of medium and stir until dissolved.
L-Cystine dihydrochloride 65.200 4. Adjust the pH to 0.2 - 0.3 pH units below the desired pH
L-Glutamic acid 20.000 using 1N HCl or 1N NaOH since the pH tends to rise during
L-Glutamine 300.000 filtration.
L-Histidine hydrochloride monohydrate 20.960 5. Make up the final volume to 1000ml with tissue culture
L-Hydroxyproline 20.000 grade water.
L-Isoleucine 50.000
Please refer disclaimer overleaf
6. Sterilize the medium immediately by filtering through a in certain instances. This can be indicated by change in
sterile membrane filter with a porosity of 0.22 micron or less, colour, change in appearance and presence of particulate
using positive pressure rather than vacuum to minimize the matter and haziness after dissolution.
loss of carbon dioxide. 2.Preparation of concentrated medium is not recommended
7. Aseptically add sterile supplements as required and since free base amino acids and salt complexes having low
dispense the desired amount of sterile medium into sterile solubility may precipitate in concentrated medium.
containers. 3.pH and sodium bicarbonate concentration of the prepared
8. Store liquid medium at 2-8°C and in dark till use. medium are critical factors affecting cell growth. This is
also influenced by amount of medium and volume of culture
Material required but not provided : vessel used (surface to volume ratio). For example, in large
bottles, such as Roux bottles pH tends to rise perceptibly as
Tissue culture grade water (TCL010)
significant volume of carbon dioxide is released. Therefore,
Sodium bicarbonate (TC230)
optimal conditions of pH, sodium bicarbonate concentration,
Sodium bicarbonate solution, 7.5% (TCL013)
surface to volume ratio must be determined for each cell type.
1N Hydrochloric acid (TCL003)
We recommend stringent monitoring of pH. If needed, pH
1N Sodium hydroxide (TCL002)
can be adjusted by using sterile 1N HCl or 1N NaOH or by
Foetal bovine serum (RM1112/RM10432)
bubbling in carbon dioxide.
4.If required, supplements can be added to the medium prior
Quality Control: to filter sterilization observing sterility precautions. Shelf life
of the medium will depend on the nature of supplement added
Appearance to the medium. L-glutamine present in the medium is a labile
Off-white to Creamish white, homogenous powder. amino acid and has a shelf life of approximately one month,
Solubility when stored at 2-8ºC.
Clear solution at 8.4 gms/L.
pH without Sodium Bicarbonate
7.20 -7.80
Revision : 1 / 2011
pH with Sodium Bicarbonate
7.40 -8.00
Osmolality without Sodium Bicarbonate
230.00 -270.00
Osmolality with Sodium Bicarbonate
280.00 -320.00
Cultural Response
The growth promotion capacity of the medium is assessed
qualitatively by analyzing the cells for the morphology and
quantitatively by estimating the cell counts and comparing it
with a control medium through minimum three subcultures.
Endotoxin Content
NMT 5EU/ml

Storage and Shelf Life:

Disclaimer :
User must ensure suitability of the product(s) in their application prior to use. Products conform solely to the information contained in this and other
related HiMedia™ publications. The information contained in this publication is based on our research and development work and is to the best
of our knowledge true and accurate. HiMedia™ Laboratories Pvt Ltd reserves the right to make changes to specifications and information related
to the products at any time. Products are not intended for human or animal diagnostic or therapeutic use but for laboratory, research or further
manufacturing use only, unless otherwise specified. Statements contained herein should not be considered as a warranty of any kind, expressed
or implied, and no liability is accepted for infringement of any patents.

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