Serum-free Medium HybriS-1
HybriS-1 serum-free medium (SFM) is used to culture of hybridomic cells lines and for synthesis of monoclonal antibodies.
The medium with alanyl-glutamine is ready to use, it was developed by PanEco. The medium is a mixture of inorganic salts, amino acids, vitamins, glucose and phenolic red dissolved in purified water, sterilized through filters with pore size of 0.22 microns. Protein-free, doesn't require serum.
A stable glutamine analogue, alanyl-glutamine, has been added to the medium, so the medium can be stored for a long time without changing the composition and doesn't require the addition of L-glutamine.
The medium is being tested at PanEco on mouse hybridoma cells.
Without Antibiotics.
This product is intended for research purposes.
- Appearance: clear, orange solution, without opalescence and sediment.
- pH: 7.0 to 7.4.
- Osmolarity: 300+20 mosmol/kg.
- Buffer capacity: at least 6.0 ml.
- The protein content: is less than 20 mg/l.
CELL ADAPTATION:
Cell lines cultured using classical (serum) media require adaptation to serum-free cultivation conditions. Adaptation period time is determined experimentally for specific cell line. Adaptation can be in 2 ways:
Direct adaptation: by simultaneous transfer of an adaptable culture into a SFM. The first passage of cells is carried out in the mid-logarithmic growth phase at concentration of 1x10⁵ cell/ml.
Sequential adaptation: by sequential passage of cells. The initial concentration in suspension is from 5x10⁴ cell/ml.
- Passage 1 serum-free media whith 2% of serum
- Passage 2 serum-free media whith 0.5% of serum
- Passage 3 serum-free media whithout serum
In each case, adaptation of the culture to SFM continues until stabilization of cell growth. Most cell lines can be considered fully adapted after 3 passages in 100% SFM. If you have trouble whith adaptation, go back and passage the cells 2–3 times in the previous ratio of serum-supplemented media to serum-free media.
Overall, cells in serum-free culture are more sensitive to extremes of pH, temperature, osmolality, mechanical forces, and enzyme treatment.
HOW SUPPLIED: in sterile plastic wide neck bottle of 500 ml.
SHELF LIFE: 1 year.
STORAGE CONDITIONS: +2 to +8 °C in a place protected from light.
RECOMMENDED USE:
- Do not use the product after its expiry date.
- Use gloves, mask and clothes adapted to the manipulation of the product to avoid contamination.
- Manipulate the product in aseptic conditions.
- Supplements such as antibiotics or L-Glutamine should be added as sterile supplements to the ready medium.
- Medium should be clear and free of opalescence and sediment.
