VP-SFM (1X)

Cells can be adapted by Sequential or Direct Adaptation. Suggested protocols for each are below, and you can also find more information by searching "Adaptation of Cell Cultures to a Serum-Free Medium" from our website home page.

SEQUENTIAL ADAPTATION
1) Subculture the cells growing in serum-supplemented medium into a 25%:75% mixture of SFM and serum supplemented medium.
2) When the cell density is 5 x 10E5 cells/ml, subculture the cells into a 50%:50% mixture of SFM and serum supplemented medium at a cell density 2.5 x 10E5 to 3 x 10E5 cells/ml.
3) Continue to subculture after the cell density 5 x 10E5 cells/ml in gradually increasing proportions of SFM until the serum is ~0.1% with about 85% cell viability.
4) Subculture the cells into SFM with an innoculum of 2.5 x 10E5 to 3 x 10E5 cells/ml.
5) When the cell density is 1 x 10E6 to 3 x 10E6 cells/ml (4 to 6 days post planting) subculture the cells again.
6) Stock cultures of SFM adapted cells should be subcultured in SFM every 3 to 5 days when the cell density is 1 x 10E6 to 3 x 10E6 cells/ml with 90% viability.

DIRECT ADAPTATION
Some cells can be directly adapted from serum-containing medium to SFM. For direct adaptation, the cell innoculum should be 1.5 x 10E5 to 3 x 10E5 cells/ml.
Cells should be subcultured when the cell density is 1 x 10E6 to 3 x 10E6 cells/ml. Cells are fully adapted to SFM when the cell density is 2 x 10E6 to 4 x 10E6 cells/ml after 4 to 7 days in culture.
Stock cultures of cells adapted to SFM should be subcultured in SFM every 3 to 5 days when the cell density is 1 x 10E6 to 3 x 10E6 cells/ml with 90% viability.

Some cells, such as insect cells, are sensitive to changes in their medium. By sequentially adapting cells, the medium is changed with minimal effects on cell growth.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Both VP-SFM (Cat. No. 11681020) and OptiPro SFM (Cat. No. 12309019) can be used to culture Vero cells and will provide comparable cell growth. VP-SFM is serum-free but not protein-free. It contains human recombinant insulin in its formulation. The human recombinant insulin uses animal origin component in its manufacturing process. OptiPRO SFM is a low protein medium that is serum-free and animal origin-free. Both products have a drug master file associated with them.

If you will be choosing either of these formulations, we would also recommend using TrypLE Express (Cat. No. 12605010 or 12604013) or TrypLE Select (Cat. No. 12563011) reagent for passaging the cells. TrypLE Express and Select reagents contain the same enzyme. The main difference is in the manufacturing process. TrypLE Select reagent is made in disposable or dedicated equipment that does not come in contact with animal origin components. TrypLE Express reagent can be manufactured in vessels that may have been used in the manufacture of cell culture media or reagents that contain animal origin components.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.