Sf9 Cells
General information
Description | 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%. |
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Organism | Fall armyworm |
Tissue | Ovary |
Applications | Transfection, plaque assay/purification, amplification of high-titer stocks, and expression of recombinant proteins |
Synonyms | SF9, sf9, SF-9, Sf-9, sf-9, Sf 9, Spodoptera frugiperda clone 9, Sf clone 9, IPLB-Sf-9AE, IPLB-SF-9AE, IPLB-SF-9, IPLB-Sf-9, IPLB-Sf9 |
Characteristics
Age | Pupal stage |
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Gender | Female |
Morphology | Round, attached, epitheloid |
Growth properties | Monolayer, adherent |
Identifiers / Biosafety / Citation
Citation | Sf9 (Cytion catalog number 604328) |
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Biosafety level | 1 |
Expression / Mutation
Virus susceptibility | Baculoviruses, Autographa californica (MNPV), St. Louis encephalitis (SLE) |
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Handling
Culture Medium | Spodopan (PAN Biotech) |
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Medium supplements | Supplement the medium with 2% FBS to enhance proliferation if needed |
Passaging solution | Accutase |
Subculturing | Detachment of cells via a cell scraper is recommended. Collect the medium with detached cells after scraping in a 15ml centrifuge tube. Add about 5ml of medium to the flask and rinse the flask several times to collect any remaining cells and combine them with the rest of the cells in the tube. Centrifuge for 3 min at 300xg, remove the supernatant, resuspend the cells in fresh, cold medium and dispense into new flasks. |
Split ratio | 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 |
Seeding density | 1 x 10^4 cells/cm^2. Incubate between 26 to 30 degree Celsius in a non-humidified, ambient air-regulated incubator. Use cell culture flasks with filter caps or loosen caps to allow for oxygen exchange. |
Fluid renewal | 2 to 3 times per week |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,x
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