| Field | Specification |
|---|---|
| Species |
- Protein expression: p53 negative
- Isoenzymes: Me-2, 1, PGM3, 1, PGM1, 2, ES-D, 1, AK-1, 1, GLO-1, 2, G6PD, B, Phenotype Frequency Product: 0.0031
- Tumorigenic: Yes, in nude mice. Forms poorly differentiated carcinoma
- Mutational profile: CaLu-6 cells carry a mutation in KRAS codon 61, c.181C>A p.(Gln61Lys). NRAS of BRAF mutation was not detected.
- Karyotype: The stemline chromosome number is hypotriploid and the 2S component occurred at 5.8%. Modal chromosome number is 59. Fourteen marker chromosomes (constitutive) were common to most S metaphases. No Y chromosome was detected in the QM stained preparation.
- cultureMedium: EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a)
- supplements: Supplement the medium with 10% FBS and 1% NEAA
- dissociationReagent: Accutase
- 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: 2 x 104 cells/cm2 will result in a 90% confluent monolayer in about 4 days
- 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 48 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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