OptiPRO™ SFM

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.

For virus titering in OptiPRO SFM we recommend using Falcon or equivalent 96-well plates and plating cells at a concentration of 1 X 10^6/mL. If you are having trouble growing cells for virus titration in OptiPRO SFM in the 96-well plates, we suggest switching to the EMEM with 5%FBS or DMEM with 5% FBS.

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

The sequential adaptation process is a multi-subculture of cells. We recommend the following passaging sequence:
• First passage: use 25% OptiPRO SFM:75% original stock medium
• Second passage: use 50% OptiPRO SFM:50% original stock medium
• Third passage: use 75% OptiPRO SFM:25% original stock medium
• Fourth and subsequent passages: use 100% OptiPRO SFM

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

You can supplement OptiPRO SFM with 5mL/L Antibiotic-Antimycotic (Cat. No. 15240096)

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