| Field | Specification |
|---|---|
| Mfr No | |
| Species |
Leukocytes and sera from patients with transitional cell carcinoma were cytotoxic to T24 and related lines.
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- Antigen expression: HLA A1, A3, B18, Bw35, Cw4, DRw2, Dw4
- Isoenzymes: Me-2, 1-2, PGM3, 1, PGM1, 1, ES-D, 1, AK-1, 1, GLO-1, 1, G6PD, B, Phenotype Frequency Product: 0.0216
- Oncogenes: H-ras+
- Tumorigenic: Yes, in hamster cheek pouch. Not in nude mice
- Products: Tumor specific antigen
- Karyotype: Hypodiploidy to hypopentaploidy, stemline 86, 2 to 4 telocentrics, 3 to 4 minutes, hypotetraploid to hypertetraploid with abnormalities including dicentrics, breaks, pulverization, minutes and telocentric markers
- cultureMedium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
- supplements: Supplement the medium with 5% FBS
- dissociationReagent: Accutase
- doublingTime: 19 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
- 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.
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