| Field | Specification |
|---|---|
| Mfr No | |
| Species |
- Antigen expression: CD326+
- Viruses: Free of human pathogenic viruses SV40, JC/BK, HBV, HCV, HIV.
- cultureMedium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 29 hours
- subculturing: Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add TrypLE Express (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at 37 degree Celsius for 10 to 15 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300xg, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium. This cell line will result in single cell suspension. It is recommended to use collagen-coated flasks.
- seedingDensity: 2 x 104 cells/cm2
- fluidRenewal: Every 3 to 5 days
- postThawRecovery: Fast
- 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.