OptiPRO™ SFM, 1000 mL - FAQs

How do I adapt my cells to serum-free medium?

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.

Why is it necessary to gradually adapt the cells to serum-free medium?

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.

Do you offer a protocol for titering virus in OptiPRO SFM?

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.

Is there a recommended protocol for culturing cell lines in OptiPRO SFM that require sequential adaptation?

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.

Can I add antibiotics to OptiPRO SFM?

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

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What concentration of L-glutamine or GlutaMAX-I should I add to OptiPRO SFM before I use it?

We recommend adding L-glutamine or GlutaMAX-I to OptiPRO SFM at a final concentration of 4mM.

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

Can you please recommend the best serum-free medium for culture of Vero cells and production of virus-like particles from these cells?

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.