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Reagents and instruments for immunology, cell biology and molecular biology.

MEL-CLS-3

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Cat# 300293

Size : 1cryovial

Contact local distributor :

Phone : +1 850 650 7790

Brand : CLS Cell Lines Service

Contact local distributor :

Phone : +1 850 650 7790

MEL-CLS-3 Cells

Description	This cell line was established as in vitro cell line from the human melanoma xenograft MRI-H-221 of a primary malignant melanoma by CLS in 1998.
Organism	Human
Tissue	Skin
Disease	Amelanotic melanoma
Synonyms	MRI-H-221

Age	Unspecified
Gender	Male
Ethnicity	Caucasian
Growth properties	Adherent

Citation	MEL-CLS-3 (Cytion catalog number 300293)
Biosafety level	1

Protein expression	p53(+)
Tumorigenic	Yes, in nude mice
Viruses	Negative for: Sendai, Ektromelie, 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)

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)
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	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Optionally, skip centrifugation but remove any remaining freezing medium after 24 hours. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

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.
STR profile	Amelogenin: x,y CSF1PO: 10,11 D13S317: 11,13 D16S539: 10,13 D5S818: 11,12 D7S820: 7,10 TH01: 9.3 TPOX: 8 vWA: 17,18 D3S1358: 16,18 D21S11: 28,31.2 D18S51: 16,17 Penta E: 14 Penta D: 12,13 D8S1179: 12,13 FGA: 21,25