CELL
CONCEPTS cell culture media raw material and components are of highest
quality. They are purchased from approved suppliers only. All chemicals
can be traced back to the lots used. During the manufacturing process,
the weights of the chemicals are controlled automatically by computerized
balances. This guarantees a high degree of reproducibility of the formula.
The water system is charakterized by resins that banish all organic and
inorganic solutes, filtration down to pore size of 0.2 µm to remove
particles and microorganisms, and finally reverse osmosis, result in ultra
pure, apyrogenic water, conform to USP standards. These rigorous purification
steps guarantee that the water used for incorporation into our cell culture
media has the utmost purity required.
The
sizes of the batches produced range from 1 to 1.200 liters, giving CELL
CONCEPTS the flexibility to better serve its customers.All
liquid media are routinely tested for sterility, growth promotion and absence
of cytotoxicity, the osmolality required and the correct pH level. Powdered
media are reconstituted with cell culture grade water and checked as described
for liquid media.
·Guide
to supplementation of media
Unless
noted otherwise, all of our media are formulated without antibiotics, serum
and L-Glutamine.
10
x concentrated media do not contain antibiotics, serum, L-Glutamine and
Sodium Bicarbonate. Powder
media contain L-Glutamine but no Sodium Bicarbonate. We
recommend to aseptically add the required amount of these supplements prior
to use.
Request more
information regarding 10x concentrates by e-mail.
Request more
information regarding powder media by e-mail
For
the addition of antibiotics, please refer to section 5, data sheet 5-011.
For
the addition of appropriate amounts of L-Glutamine (Art. No. R-L1000-H)
and Sodium Bicarbonate NaHCO3 refer to the following table 1.
Sodium Bicarbonate (Art. No. R-L3000-H) can be added either as a
solid before sterilization or as a 7.5% sterile solution after sterilization.
Serum
is usually added to the cell culture media to a final concentration of
1 to 25%.
|
Medium
(1 l)
|
L-Glutamine
(200
mM/l) mlrequired |
final
concentration of L-Glutamine (mg/l)
|
NaHCO3
(7.5%
solution) mlrequired |
NaHCO3
(solid) grequired |
final
concentration
of NaHCO3 (mg/l) |
|
BME,
EBSS
|
10.0
|
292
|
29.3
|
2.20
|
2200
|
|
CMRL-1066
|
3.4
|
100
|
29.3
|
2.20
|
2200
|
|
DMEM
|
20.0
|
584
|
49.3
|
3.70
|
3700
|
|
DMEM/Ham
F12
|
20.0
|
365
|
32.5
|
2.438
|
2438
|
|
DMEM/Ham
F12w HEPES
|
20.0
|
365
|
16.0
|
1.20
|
1200
|
|
Ham’s
F-10
|
5.0
|
146
|
16.0
|
1.20
|
1200
|
|
Ham’s
F-12
|
5.0
|
146
|
15.7
|
1.176
|
1176
|
|
MEM
Iscove
|
20.0
|
584
|
40.4
|
3.024
|
3024
|
|
Leibovitz
L-15
|
10.3
|
300
|
-
|
-
|
|
|
M199
Earle’s
|
3.4
|
100
|
29.3
|
2.20
|
2200
|
|
M199
Hank’s
|
3.4
|
100
|
4.7
|
0.35
|
350
|
|
McCoy’s
5A
|
7.5
|
219
|
29.3
|
2.20
|
2200
|
|
MEM
Alpha
|
10.0
|
292
|
29.3
|
2.20
|
2200
|
|
MEM
D-Valin
|
10.0
|
292
|
29.3
|
2.20
|
2200
|
|
MEM
Earle’s
|
10.0
|
292
|
29.3
|
2.20
|
2200
|
|
MEM
Hank’s
|
10.0
|
292
|
4.7
|
0.35
|
350
|
|
RPMI
1640
|
10.3
|
300
|
26.7
|
2.00
|
2000
|
|
Waymouth
|
12.0
|
350
|
29.9
|
2.24
|
2240
|
|
William’s
D/E
|
10.0
|
292
|
29.3
|
2.20
|
2200
|
Table
1:
Guide to addition of L-Glutamine (200 mM solution) and of Sodium Bicarbonate
(7.5% solution or powder).
1.Aseptically
measure out approximately 850 ml of sterile tissue culture grade water
in a container of appropriate size.
2.Gently
stir and aseptically add 100 ml of 10 x medium.
3.Now
add the required amount of Sodium Bicarbonate, according to table 1.
4.Sera,
antibiotics, L-Glutamine and other supplements may now be added to the
solution.
5.Use
sterile 1 N NaOH or 1 N HCl to adjust the pH of the solution to the desired
value.
6.Use
tissue grade water to adjust the final volume of the solution to 1 l.
7.The
completed medium has a limited shelf life and should be stored refrigerated.
·Preparation
of 1 l liquid media from powder media
1.Place
approximately 900 ml of cell culture water in a 1 l Erlenmeyer flask. While
gently stirring, slowly add the powder. Do not heat.
2.Add
solid Sodium Bicarbonate according to table 1. Stirr and make the solution
up with water to 1 l.
3. Control
the pH and, if necessary, adjust with 1 N HCl to approximately 0.2 pH units
below the value required after sterilization.
4.Use
0.2 µm filter to sterilize the medium.
·Media
with stable Glutamine
In
liquid solutions, L-Glutamine is not stable at 4°C or higher temperatures.
The molecule dissociates and, as a waste product of that reaction, toxic
ammonium is created. This restricts the stability of cell culture media
and as a consequence, stock solutions of L-Glutamine or solutions containing
Glutamine have to be stored frozen. Additionally, cell culture media have
to be changed frequently to ensure the availability of the amino acid.
Dipeptides
containing Glutamine are extremely well suited as a replacement of L-Glutamine.
They are stable over longer periods of time, even if stored at room temperature.
Cells may recover L-Glutamine via cleavage of the peptide bond.
CELL
CONCEPTS offers several media with stable Glutamine in the form of N-acetyl-L-Ala-L-Gln.
The amount of available L-Glutamine corresponds to that of L-Glutamine
normally present in the equivalent medium after the addition of the appropriate
amount of L-Glutamine (see table 1).
·Media
with HEPES
If
you have a sensitive cell culture system, we recommend to use of HEPES
buffered media. Such nutrient solutions are prepared in accordance with
the original formulation and include Sodium Bicarbonate. However, to maintain
an appropriate osmolality, the concentration of Sodium Chloride is decreased.
HEPES increases the buffering capacity and the stability of the pH in the
range of 7.2 to 7.6. This allows the media to better resist fluctuations
in the pH resulting from changes of cellular metabolism especially in rapidly
growing cultures with a strong acidulation.
CELL
CONCEPTS offers several media with HEPES. Contactsales@cellconcepts.de.
·Media
without Phenol Red
Phenol
Red is not a natural substance. Nevertheless Phenol Red is often added
as an indicator of the pH to cell culture media. However, the presence
of this dye is not always desired. Especially in the cultivation of primary
cells, media with Phenol Red may produce artifacts. Additionally, Phenol
Red may interfere with chromogenic bioassays.
CELL
CONCEPTS offers several media without Phenol Red.Contactsales@cellconcepts.de.
·Deficient
media
Deficient MEM media are suited for radioactive incorporation studies to investigate metabolic aspects of cellular physiology.
CELL
CONCEPTS offers several deficient MEM-media of customers inquiry. Contactsales@cellconcepts.de.
·Serum
free media
Serum
has normally to be added to cell culture media. It has multiple functions;
e.g. it may serve as a source of nutrients, enzymes, hormones and growth
factors; it may also serve as a pH-buffer or may bind toxic components.
However, serum has quite a lot of disadvantages: it is expensive, shows
lot-to-lot-variability and increases the complexity of the down-stream
processing of the desired biological material. CELL CONCEPTS offers several
serum free cell specific media media. Request
information by e-mail.
·Customer-designed
media
Do
you wish a specialised modification of a standard cell culture medium or
do you need a nutrient mixture especially adapted to the requirements of
your cells? Then CELL CONCEPTS is your appropriate partner! We produce
under strictly controlled conditions your special wish according to your
formulation. Several years of experience of our employees in the production
of cell culture media of highest quality as well as extensive in-process
end end-controls guarantee, that your product has the quality that you
expect.
If
you want to discuss additional questions to your media requirements or
to order directly your special media, please write down your formulation
and e-mail to contract.service@cellconcepts.de.
·Storage
conditions and shelf life of media
Storage
conditions and shelf life of media are indicated on the label.