| Field | Specification |
|---|---|
| Mfr No | |
| Species |
- Antigen expression: HLA A1, A10, A11, B7, B8
- Isoenzymes: G6PD, B, PGM1, 1, PGM3, 2, ES-D, 1, Me-2, 0, GLO-1, 2, AK-1, 1-2, Phenotype Frequency Product: 0.0008
- Tumorigenic: Yes, in nude mice. At passage levels below 35 the line is not tumorigenic in nude mice, but forms colonies in soft agar. Tumorigenicity has been reported to increase above passage 35.
- Viruses: The cells contain a tamdemly integrated SV40 genome it has been reported that they may contain a type D retrovirus that is similar or identical to Mason-Pfizer monkey virus (MPMV).
- Reverse transcriptase: Positive
- MSI status: Stable (MSS)
- Karyotype: The stemline chromosome number is near triploid with the modal number of 67 chromosomes, and the 2S component occurring at 0.6%. Most chromosome complements consist of about 39 normal and 28 marker chromosomes. Markers such as 2q, 11q+, 11q, t(2q.12), t(2q.5q?), t(6p?.16), 16pt and many others are common to most metaphases. Normal chromosomes 11, 14, 15 and 16 are absent. 2, 12, 17 and 19 are monosomic, and the x is disomic. DNA profiling for amelogenin, a sex-chromosome-specific PCR assay that can distinguish x chromosome-specific products from Y chromosome-specific products revealed the presence of Y chromosomes in this cell line of putative female origin. Confirmation of the general findings was accomplished by QM staining, C-banding, and FISH, with a whole chromosome paint probe to the human Y chromosome.
- 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: 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.