Leibovitz's L-15 Medium
With L-Glutamine
Without Sodium bicarbonate
Product Code: AT011
Product Description :
Leibovitz's Medium was specifically designed to grow L-Lysine hydrochloride 94.000
cells in a CO2 free atmosphere. The standard sodium L-Methionine 75.000
bicarbonate/CO2 buffering system is replaced by L-Phenylalanine 125.000
combination of free basic amino acids, phosphate buffers L-Serine 200.000
and higher levels of galactose and sodium pyruvate. As a L-Threonine 300.000
result, the medium does not require supplementation with L-Tryptophan 20.000
sodium bicarbonate and can be used under conditions of L-Tyrosine Disodium Salt 276.160
free gaseous exchange with the atmosphere. The medium L-Valine 100.000
can be used to grow human tumor cells and embryonic VITAMINS
cells and also established cell lines like HeLa and Hep-2. Choline chloride 1.000
The medium is frequently used in diagnostic virology D-Ca-Pantothenate 1.000
where tissue cell lines or strains need to be grown in Folic acid 1.000
closed systems. Leibovitz's medium obviates the need of Nicotinamide 1.000
frequent medium change. Pyridoxine hydrochloride 1.000
Riboflavin-5-phosphate, Na 0.100
AT011 is Leibovitz's Medium with L-glutamine. Users
Thiamine monophosphate 1.000
are advised to review the literature for recommendations
i-Inositol 2.000
regarding medium supplementation and physiological
OTHERS
growth requirements specific for different cell lines.
D-Galactose 900.000
Phenol red Sodium Salt 11.000
Composition : Sodium pyruvate 550.000
Ingredients mg/L
INORGANIC SALTS Directions :
Calcium chloride dihydrate 185.000 1. Suspend 14.0gms in 900ml tissue culture grade water
Magnesium chloride hexahydrate 200.000 with constant, gentle stirring until the powder is completely
Magnesium sulphate anhydrous 97.720 dissolved. Do not heat the water. Note: Presence of slight
Potassium chloride 400.000 haziness in the medium is due to inherent nature of some of
Potassium phosphate, monobasic 60.000 the ingredients in the composition. However, this will not
Sodium chloride 8000.000 affect the performance of the medium.
Sodium phosphate, dibasic anhydrous 190.120 2. Adjust the pH to 0.2-0.3 pH units below the desired pH
AMINO ACIDS using 1N HCl or 1N NaOH since the pH tends to rise during
DL-Alpha alanine 450.000 filtration.
Glycine 200.000 3. Make up the final volume to 1000ml with tissue culture
L-Arginine (free base) 500.000 grade water.
L-Asparagine 250.000 4. Sterilize the medium immediately by filtering through a
L-Cysteine (free base) 120.000 sterile membrane filter with a porosity of 0.22 micron or less,
L-Glutamine 300.000 using positive pressure rather than vacuum to minimize the
L-Histidine (free base) 250.000 loss of carbon dioxide.
L-Isoleucine 250.000
L-Leucine 125.000
Please refer disclaimer overleaf
5. Aseptically add sterile supplements as required and vessel used (surface to volume ratio). For example, in large
dispense the desired amount of sterile medium into sterile bottles, such as Roux bottles pH tends to rise perceptibly as
containers. significant volume of carbon dioxide is released. Therefore,
6. Store liquid medium at 2-8ºC and in dark till use. optimal conditions of pH, sodium bicarbonate concentration,
surface to volume ratio must be determined for each cell type.
Material required but not provided : We recommend stringent monitoring of pH. If needed, pH
can be adjusted by using sterile 1N HCl or 1N NaOH or by
Tissue culture grade water (TCL010)
bubbling in carbon dioxide.
1N Hydrochloric acid (TCL003)
4. If required, supplements can be added to the medium prior
1N Sodium hydroxide (TCL002)
to or after filter sterilization observing sterility precautions.
Foetal bovine serum (RM1112/RM10432)
Shelf life of the medium will depend on the nature of
supplement added to the medium.
Quality Control:
Appearance
Off-white to Creamish white, homogenous powder. Revision : 1 / 2011
Solubility
Clear solution at 14.0gms/L.
pH without Sodium Bicarbonate
7.70 -8.30
Osmolality without Sodium Bicarbonate
300.00 -340.00
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:
1. All the powdered media and prepared liquid culture media
should be stored at 2-8°C. Use before the expiry date. Inspite
of above recommended storage condition, certain powdered
medium may show some signs of deterioration /degradation
in certain instances. This can be indicated by change in
colour, change in appearance and presence of particulate
matter and haziness after dissolution.
2. Preparation of concentrated medium is not recommended
since free base amino acids and salt complexes having low
solubility may precipitate in concentrated medium.
3. pH and sodium bicarbonate concentration of the prepared
medium are critical factors affecting cell growth. This is
also influenced by amount of medium and volume of culture
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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