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K0200-05 Keratinocyte Basal Medium MCDB 153 w/Spermidine, Low Sodium Chloride, w/o Nickel Chloride•6H2O, Putrescine•2HCl, Phenol Red, Choline Chloride (Powder)

Specifications
References
Grade
Cell Culture Grade
EU Commodity Code
38210000
Shipping Temp
RT
Storage Temp
RT/4°C
Base powder without supplements

MCDB media were designed for the low-protein or serum-free growth of specific cell types using hormones, growth factors, trace elements or low levels of dialyzed fetal bovine serum protein (FBSP). Each MCDB medium was formulated (qualitatively and quantitatively) to provide a defined and optimally balanced nutritional environment that selectively promoted growth of a specific cell type. MCDB 105 and 110 are modifications of MCDB 104 medium, optimized for long-term survival and rapid clonal growth of human diploid fibroblast-like cells (WI-38, MRC-5, IMR-90) and of low-passage human foreskin fibroblasts using FBSP or hormone and growth factor supplements. MCDB 151, 153, 201 and 302 are modifications of Ham's nutrient mixture F-12, designed for the growth of human keratinocytes, clonal growth of chicken embryo fibroblasts and chinese hamster ovary (CHO) cells using low levels of FBSP, extensive trace elements or no serum protein. The selection of a nutrient medium is strongly influenced by 1. type of cell; 2. type of culture (monolayer, suspension, clonal); and 3. degree of chemical definition necessary. It is important to review the literature for recommendations concerning medium, supplementation and physiological parameters required for a specific cell line. MCDB 153 is a basal medium for the growth of normal keratinocytes and squamous epithelial cells after growth factor supplementation. Calcium content is low. Supplement with growth factors or fetal calf serum.

Appearance
White to off-white, homogeneous, free flowing powder
Solubility
Colorless, clear, complete
pH
As Reported
Endotoxin
≤1EU/ml
Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Directions per Liter
Dissolve 10.30g in 800-900ml of ddH2O stirring gently until completely solubilized. Add 1.2g/L sodium bicarbonate. Adjust pH of the medium to the desired level. Add additional water to bring the solution to 1L. Filter-sterilize using a 0.22 micron membrane filter. Aliquot into sterile containers. Do not autoclave. Contains heat-labile compounds that can be damaged with autoclaving.
Storage and Stability
Store powdered media at RT. Stable for 12 months after receipt. Opened bottles should be capped tightly and kept in a dark, low humidity environment. Prepared media should be kept at 4°C and used within a short period of time.
Media Formulation
Components shown as mg/liter
Inorganic Salts
Ammonium Metavandate0.000585
Ammonium Molybdate•4H2O0.0012359
Calcium Chloride•2H2O3.33
Cupric Sulfate•5H2O0.0027467
Ferric Nitrate•9H2O1.39
Magnesium Chloride57.13
Manganese Sulfate•5H2O0.00241
Nickel Chloride•6H2OAbsent
Potassium Chloride111.83
Sodium Acetate303.4
Sodium Chloride379.6
Sodium Metasilicate•9H2O0.1421
Sodium Phosphate Monobasic284.2
Sodium Pyruvate55.0
Sodium Selenite•5H2O0.00789
Stannous Chloride•2H2O0.0001128
Zinc Sulfate•7H2O0.14375
Amino Acids
L-Alanine8.9
L-Arginine174.2
L-Asparagine13.21
L-Aspartic Acid3.993
L-Cysteine29.33
L-Glutamic Acid14.71
L-Glutamine876.0
Glycine7.5
L-Histidine12.416
L-Isoleucine1.97
L-Leucine65.6
L-Lysine14.62
L-Methionine4.48
L-Phenylalanine4.956
L-Proline34.53
L-Serine63.03
L-Threonine11.91
L-Tryptophan3.063
L-Tyrosine2.718
L-Valine35.16
Vitamins
Biotin0.012658
Choline ChlorideAbsent
Vitamin B120.40662
Folic Acid0.79452
Myo-Inositol18.02
Nicotinamide0.03663
D-Pantothenic Acid•Ca0.2383
Pyridoxine•HCl0.02056
Riboflavin0.03764
Thiamine•HCl0.3373
Other
Adenine24.318
Thymidine0.7266
D-Glucose1080.2
HEPES Free Acid6600.0
Lipoic Acid0.2063
Phenol Red, SodiumAbsent
Putrescine•2HClAbsent
Spermidine0.3222
Total:10.30g/liter
References
1. Boyce, S.T.,Ham, R.G.: Calcium-Regulated Differentiation of Normal Human Epidermal Keratinocytes in Chemically Defined Clonal Culture and Serum-Free Serial Culture. J. Invest. Dermatol. 81: 33-40 (1983). 2. McKeehan, W.L., Ham, R.G.: Stimulation of Clonal Growth of Normal Fibroblasts with Substrata Coated with Basic Polymers. J. Cell Biol. 71: 727-734 (1976). 3. Hamilton, W.G., Ham, R.G.: Clonal Growth of Chinese Hamster Ovary Cell Lines in Protein-Free Media. In Vitro 13(9): 537-547 (1977).
USBio References
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