SF-9 Cell Growth Medium, Protein-free medium, w: L-Glutamine, w: 0.35 g/L NaHCO3

SF-9 Cell Growth Medium is a serum- and protein-free medium optimized for the growth of insect cells like Sf9 and Sf21 (Spodoptera frugiperda). SF-9 Cell Growth Medium has been developed for the cultivation of insect cells and is suitable for the production of recombinant proteins (Baculovirus expression vector system).

Usage instructions

You can follow this protocol to adapt your insect cell line to protein-free culture in a serum-free medium, for SF-9 cells. The optimal temperature range for most insect cells is 25°C to 30°C (27°C incubation +/- 0.5°C). The pH for cell culture with Sf9, and Sf21 should be between pH 6.0 and pH 6.4, and the osmolality should be 345-380 mOsm/kg. For oxygen supply, use vessels with filter lids or slightly unscrew the lids. For adaption from serum-containing to protein-free culture, direct or sequential adaption can be done. Cells should be taken from the middle exponential growth phase with >90% viability.

Direct adaptation

• Transfer cells from serum-containing culture directly into SF-9 Cell Growth Medium (prewarmed to ambient temperature) with a density of 0.5 million cells/ml.
• Subculture cells at > 2 million cells/ml, seed into fresh SF-9 Cell Growth Medium with a density of 0.5 million cells/ml.
• Repeat the subculture routine till the cells reach > 80% viability.

Indirect adaptation

• Start with subculturing the cells into a 1+1 mixture of the serum-containing medium and serum-free medium, for SF-9 cells, with a density of 0.5 million cells/ml.
• At > 1 million cells/ml, dilute to 0.5 million cells/ml by adding the respective volume of serum-free medium, for SF-9 cells.
• Repeat this process till serum levels below 0.1% is reached; the viability should be above 80%. Cell density should exceed 1 million cells/ml.

Quality control

• pH = 7.2 +/- 0.02 at 20-25°C
• Each lot has been tested for sterility and absence of mycoplasma and bacteria

Maintenance

• Keep refrigerated at +2°C to +8°C in the dark. Freezing as well as warming up to +37°C minimizes the quality of the product.
• Do not heat the medium to more than 37°C or use uncontrollable sources of heat (e.g., microwave appliances).
• If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
• Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.

Ingredients

SF-9 Cell Growth Medium contains amino acids, vitamins, salts, trace elements, lipids, and growth-promoting factors in a formulation optimized for insect cells. It is free of proteins and any components of human origin.

Disclaimer

This product is for in-vitro use only. Not intended for clinical or diagnostic applications.