| Field | Specification |
|---|---|
| Mfr No | |
| Species |
- Receptors expressed: Epidermal growth factor (EGF), keratin (immunoperoxidase staining). Matrilysin, a metalloproteinase associated with tumor invasiveness, is not expressed.
- Protein expression: The cells express elevated levels of p53 protein.
- Antigen expression: HLA A2, B8, B17, blood type A, Rh+. The line is negative for CSAp (CSAp-) and colon antigen 3
- Isoenzymes: G6PD, B, PGM1, 2, PGM3, 1, 6PGD, A, PEP-D, 1, ES-D, 1
- Tumorigenic: Yes, in nude mice
- Viruses: Reverse transcriptase negative
- Virus susceptibility: Human immunodeficiency virus (HIV, LAV)
- Products: Carcinoembryonic antigen (CEA) 0.7 ng/106 cells/10 days, keratin, TGF-β. The cells have been reported to produce GM-CSF.
- Mutational profile: SW-480 cells carry a homozygous Kras mutation in codon 12: GGT(Wt Gly) >GTT(Val). There is a G->A mutation in codon 273 of the p53 gene resulting in an Arg->His substitution and a C->T mutation in codon 309 resulting in a Pro->Ser substitution.
- cultureMedium: Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 20 to 25 hours
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2
- fluidRenewal: 1 to 2 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 hours.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.