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Catalog Number: M-154-500
Medium 154 is a sterile, liquid tissue culture medium prepared
with 200 µM calcium chloride for the growth of normal human
epidermal keratinocytes. This medium is intended for use as
one component in a complete culture environment. Medium
154 is a basal medium containing essential and non-essential
amino acids, vitamins, other organic compounds, trace
minerals, and inorganic salts. This medium does not contain
antibiotics, antimycotics, hormones, growth factors, or proteins.
This medium is HEPES and bicarbonate buffered and is
designed for use in an incubator with an atmosphere of 5%
normal human keratinocytes, Medium 154 must be
supplemented with Human Keratinocyte Growth Supplement
(HKGS, Cat. S-001-5) or Human Keratinocyte Growth
Supplement Kit (HKGS Kit, Cat. S-001-K).
Medium 154 is intended for use in the routine culture of normal
human epidermal keratinocytes. When supplemented with
HKGS or HKGS Kit, Medium 154 will support the plating and
proliferation of keratinocytes at varying culture densities from
clonal (25 cells/cm
) to high density (8 x 10
keratinocytes from skin, and clonal growth assays. For optimal
results when performing primary isolations, Medium 154
should be used in conjunction with Coating Matrix Kit (Cat. R-
This product is for research use only. Not for use in
Medium 154 is stored at 4° C in our facility and is shipped at
ambient temperature. Upon receipt, the medium should be
stored at 4° C and should not be frozen. Protect from light.
Several components of this tissue culture medium are light-
labile, and we recommend that the medium not be exposed to
light for lengthy periods of time. If the medium is warmed prior
to use, do not exceed 37° C. When stored in the dark at 4° C,
the product is stable until the expiration date on the label.
Please use the supplementation instructions (page 2) to
prepare the medium for use.
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Preparation of Supplemented Medium 154
1. Thaw the frozen components of the HKGS Kit, or one
bottle of HKGS according to the instructions provided with
those products. Make sure that the caps of all of the
bottles are tight. Gently swirl the bottle(s) of supplement.
Avoid splashing the supplement into the cap of the bottle
or causing the supplement to foam.
2. Wipe the outside of the containers with a disinfecting
solution such as 70% ethanol or isopropanol.
3. To add the HKGS, transfer the entire contents of the bottle
of supplement to the bottle of medium using sterile
technique in a laminar flow culture hood. To add the
HKGS Kit, transfer the desired amount of each component
of the HKGS Kit to the bottle of medium using sterile
technique in a laminar flow culture hood. Note: addition of
less than the entire amount of any component may affect
the performance of the supplemented medium. If
the same technique as above.
4. Tightly cap the bottle of supplemented medium and swirl
the contents to ensure a homogeneous solution. Avoid
causing the medium to foam.
Storage and Stability of Supplemented Medium 154
Once Medium 154 has been supplemented with HKGS or
HKGS Kit, the supplemented medium should be stored in the
dark at 4° C and should not be frozen. When stored in the
dark at 4° C, the supplemented medium is stable for 1 month.
The Medium 154 formulation is based on medium MCDB 151, with
trace elements as in medium MCDB 104, and the high amino acid
modifications of Pittelkow.
Cook, Pittelkow, and Shipley; J. Cell. Physiol. 146:277-289, 1991
Peehl and Ham; In Vitro 16: 526-540, 1980
McKeehan et al.; In Vitro 13: 399-416, 1977
Pittelkow and Scott; Mayo Clin. Proc. 61: 771-777, 1986
Cascade Biologics, Inc. (hereinafter, CBI) warrants that its products will perform according to the information provided in various publications that it
distributes, and as described herein for the intended shelf life of the product when stored under the conditions prescribed by CBI. If any product does
not perform according to the published information provided by us, CBI will replace the product free of charge to the original customer. The remedy of
product replacement shall be the customers' sole and exclusive remedy for defective product, unless CBI is unable to deliver replacement product, in
which case CBI shall reimburse the customer for the purchase price of the defective product. Customer understands that the foregoing limited
warranty is in lieu of all other warranties and CBI hereby disclaims all other warranties including, but not limited to, implied warranty of merchantability
or fitness for adequacy for any particular purpose or use. CBI shall not be liable to customer or to any party claiming through customer for any
incidental or consequential damages, including, but not limited to any lost profits, lost savings, or lost business, whether arising out of contract, tort, or
otherwise. Customer acknowledges that products are intended for research use only and are not to be used in animals, humans, or diagnostic