Serum-free medium Hybris-1-CHO
The Hybris-1-CHO serum-free medium (SFM) is intended for suspension cultivation and production of recombinant proteins by cells of genetically engineered CHO cell lines.
The product is developed by PanEco Ltd. The medium is is a mixture of inorganic salts, amino acids, vitamins, glucose and phenolic red dissolved in purified water, sterilized through filters with pore size of 0.2 microns PES filters. The medium has chemically defined composition and does not contain products of animal or plant origin.
The basic formulation of the medium does not include sources of glutamine and nucleotides. To ensure the growth of uncomplemented DHFR- or GS-mutants, it is necessary to add nucleotide precursors in the form of Hypoxanthine/Thymidine (HT) and/or Glutamine (Alanyl-Glutamine).
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 less than 10 mg/l.
PREPARATION INSTRUCTIONS:
- Open vials with the medium in sterile conditions.
- Before use, it is necessary to add a source of glutamine (alanyl-glutamine is recommended) in concentration of 4-6 mM.
- To accelerate processes of biomass accumulation and initiation of synthesis of the target product, the medium is enriched with Hypoxanthine-Thymidine solution (Cat. No Ф050 by PanEco) to the final concentration of 1.0-1.5 x.
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.
