| Field | Specification |
|---|---|
| Species |
- Protein expression: CSAp- (Centriole and Spindle-Associated protein)
- Antigen expression: The cells are positive for keratin by immunoperoxidase staining.
- Isoenzymes: G6PD, B, PGM1, 1-2, PGM3, 1-2, 6PGD, A, ES-D, 1-2, PEP-D, 1
- Tumorigenic: Yes, in nude mice
- Reverse transcriptase: Negative
- Products: Carcinoembryonic antigen (CEA) 1.5 to 4.1 ng/106 cells/10 days, keratin, interleukin 10 (IL-10, interleukin-10)
- MSI status: Stable (MSS)
- cultureMedium: RPMI 1640, w: 2.0 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a)
- supplements: Supplement the medium with 10% FBS
- doublingTime: 20 to 25 hours
- subculturing: Collect suspension cells in a 15 ml tube and carefully rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 10 minutes, then centrifuge the cells growing in suspension and the adherent cells together. Carefully resuspend the cells and dispense into new flasks which contain fresh medium.
- seedingDensity: 1 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 24 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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