| Field | Specification |
|---|---|
| Species |
- Isoenzymes: PGM3, 1-2, PGM1, 1, ES-D, 1, AK-1, 1-2, GLO-1, 2, G6PD, B,
- Tumorigenic: Yes, in nude mice. Produces undifferentiated malignant tumor, also at low frequency (22%) in the cheek pouch of cortisone treated hamsters
- 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: 45 to 50 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: An initial seeding density of 3 to 4 x 104 cells/cm2 is recommended. Later on, 2 x 104 cells/ cm2 will yield a confluent layer in 4 to 5 days.
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Within 24 to 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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