| Field | Specification |
|---|---|
| Mfr No | |
| Species |
- Isoenzymes: G6PD, B, PGM1, 1-2, PGM3, 1, AK-1, 1-2, GLO-1, 2, Phenotype Frequency Product: 0.0055
- Tumorigenic: Yes, produces tumors in nude mice consistent with fibrosarcoma
- Karyotype: Hypertriploid. Modal number = 73, range = 59 to 79. The rate of higher ploidies was 9.1%. Eleven markers were common to most cells. These include: der(2)t(2,6)(p13,q13), der(12)t(8,12)(q11,q24), t(15q21q), 19q+, t(8p21q?) and six others. Of these, the der(2) and t(8p21q?) were generally paired. A few cells had double minutes (DM)(one per cell when present).There were 4 copies of N1, N18, N20 and N22 in most cells. Normal 15 and Y were absent. The x was paired in all cells.
- 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.
- fluidRenewal: 2 to 3 times per week
- 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.