| Field | Specification |
|---|---|
| Species |
- Protein expression: p53 negative, CEA positive
- Antigen expression: Blood Type B, Rh+
- Isoenzymes: PGM3, 1, PGM1, 1, ES-D, 1, AK-1, 1, GLO-1, 2, G6PD, B, Phenotype Frequency Product: 0.0742
- Tumorigenic: Yes, in cheek pouches of anti thymocyte serum treated hamsters, not tumorigenic in nude mice
- Karyotype: The stemline chromosome number is hypotetraploid with the 2S component occurring at 6.2%. Nine markers were common to most S metaphases, four markers were less frequent. One (occasionally 2 copies) homogenous staining region (HSR) (t(11,HSR) was present in all metaphases examined, but no double minutes (DM) were detected (Sekiguchi 1978).
- cultureMedium: Ham's F12, w: 1.0 mM stable Glutamine, w: 1.0 mM Sodium pyruvate, w: 1.1 g/L NaHCO3 (Cytion article number 820600a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- doublingTime: 36 hours
- subculturing: Gather the suspension cells in a 15 ml tube and gently wash the adherent cells with PBS lacking calcium and magnesium (use 3-5 ml for T25 flasks and 5-10 ml for T75 flasks). Apply Accutase (1-2 ml for T25 flasks, 2.5 ml for T75 flasks) ensuring full coverage of the cell layer. Allow the cells to incubate at room temperature for 10 minutes. Following incubation, combine and centrifuge both the suspension and adherent cells. After centrifugation, carefully resuspend the cell pellet and transfer the cell suspension into new flasks containing fresh medium.
- seedingDensity: 2 x 104 cells/cm2 will result in a confluent monolayer within 2 to 3 days.
- fluidRenewal: Every 3 to 5 days
- 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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