Complete Culture Media

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Chromosome Medium - Liquid

The complete medium is standardized for chromosome analysis with excellent lot to lot consistancy.
INSTRUCTIONS FOR CHROMOSOME ANALYSIS

Blood samples should be taken (sufficient cannular diameter, reduced aspiration), as to avoid haemolysis.

1. Under appropriate (sterile) conditions, approx. 0.5 ml (25-30 drops) of heparinized venous whole blood are added and carefully mixed with 8 ml of Chromosome medium B (or 8 ml of Chromosome medium A + 0.08 ml PHA L (cat.no. M 5030) in a tube. Cultivation time should be 72 hrs at +35 - +37°C.

2. 0.4 ml of Colchicin/Colcemid (10 µg/ml; cat.no. L 6211/L 6221) are added (corresponding to 0.48 µg/ml of medium concentration); incubate for an additional 2 hrs.

3. Cells are centrifuged for 5 min at approx. 1000 rpm. Remove supernatant and resuspend cells in 5 ml of hypotonic potassium chloride (0.075 M; cat.no. L 6413). Incubate for another 12 min at +35 - +37°C and centrifuge the culture at 1000 rpm for 10 min.

4. Discard all but 0.25 ml of the supernatant, resuspend cells in the remaining fluid.

5. Add 5 ml of freshly prepared, ice cold fixative (glacial acetic acid/methanol 1:3) dropwise to the cell suspension while gently moving the flask. Allow resting for 10 min at +2 - +8°C.

6. Centrifuge at 1000 G for 10 min, remove remaining fixative and resuspend cells in 5 ml ice-cold fixative.

7. Following another centrifugation, cells will become visible as a slightly cloudy suspension (approx. 0.5 - 1 ml). Drop 3 - 5 drops of cell suspension from a 30 cm distance into a moist, non-greasy slide.

8. Air-dried slides can be stained readily using the Orcein or Giemsa staining technique.

Formulation(external links)

Pack/Density

Unit

Cat.no.


Chromosome Medium A without phytohemagglutinin (PHA) L Storage: 36 months at -20°C	- / -	100 ml	F 5013



Chromosome Medium B with phytohemagglutinin (PHA) L Storage: 36 months at -20°C	- / -	100 ml	F 5023

Hybridoma Medium - Liquid

Optimized for the production of monoclonal antibodies.
Different from the original formulation, only 0.05 mg/l riboflavin are used due to danger of photo oxidation.

Pack/Density

Unit

Cat.no.


Hybridoma medium Formulation (external links) without HAT with L-glutamine with 10% FBS	- / -	500 ml	F 5515

HybridoMed DIF 1000 - Liquid

Serum free medium for mammalian cells

The medium was developed to effectively grow hybridomas, but is also suitable for other cell lines. It is based on a 1:1 mixture of Iscove's medium and Ham's F-12, supplemented with transferrin, insulin, and a BSA/oleic acid complex. Myelomas, hybridomas and also other lymphoic and non-lymphoic cell lines are cultivated in Hybridomed and show growth rates comparable to those observed in serum-supplemented media. Successfully cultivated cells include: YAC-1 (mouse T-cell lymphoma), HeLa (human epitheloid cancinoma), BJA-B (human EBV-negative myeloma), BHK 21 (syrian hamster kidney) and L-psv 129 (mouse L-fibroblast). The medium is available for both 5%, and 10% CO2 atmosphere. For use in bioreactors or to cultivate delicate normal cells, we recommend cat.no. F 8035 (CO2 should be 10% to reach physiological pH values). With incubators adjusted at 5% CO2, F 8055 is recommended. This medium has a reduced level of NaHCO3 and contains 3 g/l HEPES additionally.
Different from the original formulation, only 0.05 mg/l riboflavin is used due to risk of photo oxidative effects
A trypsin inhibitor (cat.no. L 2183) is required with serum free media after trypsinating!

Formulation (external links)

Pack/Density

Unit

Cat.no.


HybridoMed DIF 1000 liquid with L-glutamine with NaHCO₃ for 10% CO₂ Storage: 36 months at -20°C	- / -	500 ml