| Field | Specification |
|---|---|
| Species |
- Receptors expressed: The cells express the wildtype and variant estrogen receptors as well as progesterone receptor.
- Protein expression: p53 negative, pGP9.5 negative, CEA positive
- Isoenzymes: PGM3, 1, PGM1, 1-2, ES-D, 1-2, AK-1, 1, GLO-1, 1-2, G6PD, B,
- Oncogenes: wnt7h +, Tx-4
- Tumorigenic: Yes, in nude mice
- Products: Insulin-like growth factor binding proteins (IGFBP) BP-2, BP-4, BP-5
- Mutational profile: TP53 wt
- Karyotype: The stemline chromosome numbers ranged from hypertriploidy to hypotetraploidy, with the 2S component occurring at 1%. There were 29 to 34 marker chromosomes per S metaphase, 24 to 28 markers occurred in at least 30% of cells, and generally one large submetacentric (M1) and 3 large subtelocentric (M2, M3, and M4) markers were recognizable in over 80% of metaphases. No DM were detected. Chromosome 20 was nullisomic and x was disomic. Phenotype Frequency Product: 0.0154
- 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
- doublingTime: 24 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: 3 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Allow the cells to rest for 48 hours past thawing
- 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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