Serum-free medium HybriS-2
The serum-free medium Hybris-2 is used for culture of myeloma and mouse hybridoma cells to produce monoclonal antibodies, and is able to maintain sufficient culture density with high antibody productivity for 2-3 passages. With addition of 1-2% ECS, it can be used for long-term cell cultivation with the same efficiency as a classic medium which contains 10% of ECS.
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. The protein content in HybriS-2 medium, due to the addition of bovine serum albumin, is no more than 550 mg/l.
Without Antibiotics.
This product is intended for research purposes.
- Appearance: clear, yellow solution, without opalescence and sediment.
- pH: 7.0 to 7.4.
- Osmolarity: 320+20 mosmol/kg.
- Buffer capacity: at least 6.0 ml.
- The protein content: is no more than 550 mg/l.
PREPARATION INSTRUCTIONS:
- Before use, add 146 mg of dry sterile L-glutamine to a bottle with medium (500 ml). Manipulate the product in aseptic conditions.
- If it necessary, antibiotics and antimycotics should be added to the medium.
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 2-5x10⁵ 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.
