| Field | Specification |
|---|---|
| Species |
- Protein expression: p53 positive
- Antigen expression: Blood Type A, Rh+, HLA A11, Bw22(+/-), B40, B18
- Isoenzymes: PGM3, 1, PGM1, 1-2, ES-D, 1, AK-1, 1-2, GLO-1, 2, G6PD, B, Phenotype Frequency Product: 0.0044
- Tumorigenic: Yes, in nude mice, forms poorly differentiated adenocarcinoma
- Mutational profile: TP53 mut
- Karyotype: (P9) hypertriploid to hypotetraploid (+A, +B, +C, +E, +F, +G, -D) with abnormalities including dicentrics, acrocentric fragments, rings, secondary constrictions, large metacentrics or polycentrics and large submetacentric marker
- cultureMedium: McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 30 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: Start culture from cryovial at 3 x 104 cells/cm2. Use 2 x 104 cells/cm2 for continued subcultures
- fluidRenewal: 2 to 3 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.
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