Sf9 Cells

Transfection, plaque assay/purification, amplification of high-titer stocks, and expression of recombinant proteins

Sf9 cells are clonal isolates derived from the Spodoptera frugiperda Sf21 cell line (IPLB-Sf-21-AE). They are commonly used in insect cell culture for recombinant protein production using baculovirus expression systems. Sf9 cells are epithelial in morphology and were cloned from the pupal ovarian tissue of the fall armyworm.
One of the key characteristics of Sf9 cells is their small, regular size which is ideal for the formation of monolayers and plaques. They are also suitable for transfection, plaque assay/purification, amplification of high-titer stocks, and expression of recombinant proteins. The Sf9 insect cell line can be maintained in attached and suspended cultures, and do not require serum or CO2 to grow.
They are considered Biosafety Level 1 and are usually grown in a 26-28 degree celsius incubator. Sf9 cells/baculovirus expression systems are widely used for high-level protein expression, often for purification, but proteins may also be functionally expressed in the defined Sf9 cell environment. The size of infected Sf9 cells is generally 17-30 microns in diameter.
The Sf9 cell line is distinct from the Sf21 cell line in that it is a clonal isolate with a smaller and more regular size, while Sf21 cells are more disparate in size and form monolayers and plaques that are more irregular.
Some Sf9 cell lines may harbor a negative sense Rhabdovirus called Spodoptera frugiperda rhabdovirus (SfRV), although not all tested Sf9 cells appear to be infected with this virus. The genome size of Sf9 has been estimated to be 451 Mbp with a G+C content of 36.53%.

Monolayer, adherent

Female

1

Supplement the medium with 2% FBS to enhance proliferation if needed

2 to 3 times per week

Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

Upon receipt, either store the cryovial immediately at temperatures below -150° C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

For immediate culturing, swiftly thaw the vial by immersing it in a 37° C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.

We stand by the promise of delivering products with high cell viability and robust culture performance. To achieve the best results, please make sure you follow the storage and culture instructions detailed in the product information sheet closely. Your adherence to these guidelines is key to success.

For the first two subcultivations a ratio of 1:3 to 1:5 is recommended. In further subcultivations cells can be split at a ratio of 1:10 to 1:20

Baculoviruses, Autographa californica (MNPV), St. Louis encephalitis (SLE)