MEL-CLS-4 growing culture
Référence 330128
Conditionnement : 1cellcultureflask
Marque : CLS Cell Lines Service
MEL-CLS-4 Cells
General information
Description | This cell line was established as in vitro cell line from the human Melanoma xenograft BOW-G by CLS in 1998. |
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Organism | Human |
Tissue | Skin |
Disease | Melanosarcoma |
Characteristics
Age | Unspecified |
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Gender | Male |
Ethnicity | Caucasian |
Growth properties | Adherent |
Identifiers / Biosafety / Citation
Citation | MEL-CLS-4 (Cytion catalog number 300128) |
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Biosafety level | 1 |
Expression / Mutation
Tumorigenic | Yes, in nude mice |
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Viruses | Negative for: Sendai, Ektromelia, Polyoma, K-Virus, Kilham, Reo 3, PVM, LCM, M.pulmonis, MVM, Theiler's GD VII, Toolan's H-1, MHV, LDV, RCV/SDA, M-Adenovirus, B.piliformis. |
Mutational profile | V600E type BRAF Mutation was determined by DNA based methods (sequencing, RT-PCR) and protein based methods (Western Blot) |
Handling
Culture Medium | DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | 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. |
Split ratio | A ratio of 1:2 to 1:4 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | Every 3 days |
Freezing recovery | After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
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Quality control / Genetic profile / HLA
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile | CSF1PO: 11,12 D13S317: 12 D16S539: 13 D5S818: 13 D7S820: 10,11 TH01: 6.9 TPOX: 8,11 vWA: 17,18 D3S1358: 15,18 D21S11: 30 D18S51: 11,14 Penta E: 7,19 Penta D: 13 D8S1179: 13,16 FGA: 19,23 |