· Technical Information

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). 

· Preparation of 1 l single strength media from 10 x concentrate

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.