| Field | Specification |
|---|---|
| Species |
- Receptors expressed: urokinase receptor(u-PAR), vitamin D (moderate expression), no detectable plasminogen activator activity.
- Protein expression: CEA negative, p53 positive
- Antigen expression: Blood Type A, Rh+, HLA A1, A3, B12, B17, Cw5, CD4 -, cell surface expression of galactose ceramide (a possible alternative receptor for HIV)
- Isoenzymes: Me-2, 1, PGM3, 1-2, PGM1, 1-2, ES-D, 1, AK-1, 1, GLO-1, 1-2, G6PD, B, Phenotype Frequency Product: 0.0230
- Oncogenes: myc+, ras+, myb+, fos+, sis+, p53+, abl -, ros -, src -
- Tumorigenic: Yes, in nude mice. Forms well differentiated adenocarcinoma consistent with colonic primary (grade I), tumors also form in steroid treated hamsters
- Virus susceptibility: human immunodeficiency virus (HIV, LAV)
- Products: Secretory component of IgA, carcinoembryonic antigen (CEA), transforming growth factor beta binding protein, mucin, The p53 antigen is overproduced
- Karyotype: The stemline chromosome number is hypertriploid with the 2S component occurring at 2.4%. Seventeen marker chromosomes are found in most metaphases, generally in single copy per chromosome. The marker designations are: M1p-(=t(3p-,?) with a deleted short arm), t(7q,?), t(10q,?), i(13q), 19q+a. M6, ?t(8q,9q-), ?xp, M9, 6q+, t(13,?)a, t(13,?)b, 19q+b, M14, M15, 15p+, and xq-. Chromosome 13 is nullisomic and chromosomes 8 and 14 are generally monosomic. No Y chromosome was detected by QM band analysis.
- 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: 24 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: 3 x 104 cells/cm2
- fluidRenewal: 2 to 3 times per week
- postThawRecovery: Slow, the cells need roughly 48 hours to settle and adhere.
- 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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