| Field | Specification |
|---|---|
| Species |
- Antigen expression: Blood Type A, Rh+, HLA Aw24, A28, B12, Bw47
- Isoenzymes: Me-2, 1, PGM3, 2, PGM1, 2, ES-D, 1, AK-1, 1-2, GLO-1, 1-2, G6PD, B, Phenotype Frequency Product: 0.0001
- Tumorigenic: Yes, in nude mice
- Karyotype: The line has a near pentaploid chromosome number and a wide range of chromosome number distribution (40% of the cells had numbers ranging from 110 to 115). The following 14 markers were found in most metaphases: t(1p,2p), t(3p,?), t(4p,11q), t(7p,22q), M6, t(9q,?), i(11q)18q t(10q,?), M14, M15, M16, M17 and t(10q,22q), 6 of these were found in some and 10 were seen in one only. Normal chromosomes 7, 8, 12, 19, 20 and 22 had 5 to 6 copies per cell, the x had two copies and the Y was absent.
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- 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
- fluidRenewal: 2 to 3 times per week
- freezeMedium: As a cryopreservation medium, use 50% basal medium + 40% FBS + 10% DMSO, 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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