| Field | Specification |
|---|---|
| Species |
- Antigen expression: CDx (+/-), CDy (-),
- Isoenzymes: AK-1, 1, ES-D, 1-2, GLO-1, 2, G6PD, B, PGM1, 1, PGM3, 1, Me-2, 1
- Tumorigenic: In nude mice
- Viruses: Reverse Transcriptase negative
- Products: Carcinoembryonic antigen (CEA) 0.5 ng/10 exp6 cells/10 days, alkaline phosphatase, keratin
- Mutational profile: HRT-18 cells carry a mutation in codon 13 of Kras gene: GGC(Wt Gly) >GAC(Asp)
- 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 10% FBS
- dissociationReagent: Accutase
- doublingTime: 15 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: 2 to 4 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Fast
- 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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