| Field | Specification |
|---|---|
| Mfr No | |
| Species |
Protein expression: DP (desmoplakin) +, PG (Plakoglobin) +, PP1 -, PP2 +, PP3 - (PP=Plakophilin), Dsc1 -, Dsc2 +, Dsc3 + (Dsc=Desmocollin), Dsg1 -, Dsg2 +, Dsg3 - (Dsg=Desmoglein), N-Cadherin +, PGP2 +.
- cultureMedium: McCoys 5a, w: 3.0 g/L Glucose, w: stable Glutamine, w: 2.0 mM Sodium pyruvate, w: 2.2 g/L NaHCO3 (Cytion article number 820200a)
- 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: 5 x 103 cells/cm2 will yield in a confluent layer in about 4 days. Seeding densities of more than 9x 103 cells/cm2 are not recommended
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Allow the cells to adhere for at least 24 to 48 hours.
- 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.