Melanoma cell lines WM3734, 1205 LU, Mel1617 and 451 LU were kindly gifted by M. Herlyn from the Wistar Institute (Philadelphia, USA)20 (link). A375, SK-MEL 19 and SK-MEL 28 cell lines were purchased from ATCC. All melanoma cells used were BRAFV600E-mutated metastatic melanoma cell lines and exhibit different TP53 gene mutational status. According to the categories and data previously described and available at IARC TP53 data base21 (link), A375, WM3734, 1205Lu and Mel1617 are TP53 wild-type cell lines, TP53 mutation of the SK-MEL 28 (TP53 L145R) and 451Lu (TP53 Y220C) cell line leads to the expression of a non-functional p53 protein and the TP53 mutation of the SK-MEL 19 cell line (TP53 N131K) still enables partially functional p53 protein expression. Melanoma cell lines were cultured as described previously22 (link). MAPKi-resistant melanoma cell lines were generated by treatment with continuously increasing concentration of the BRAF inhibitor (BRAFi) vemurafenib (up to 2 µM) or additionally with the MEK inhibitor (MEKi) trametinib (up to 50 nM) for several months.
Sk mel 19
Manufactured by American Type Culture Collection
Sourced in China, Spain
SK-MEL-19 is a human melanoma cell line derived from the skin of a 51-year-old male patient. This cell line exhibits epithelial-like morphology and is commonly used in cancer research and drug development studies.
Automatically generated - may contain errors
Lab products found in correlation
Sk mel 28, by American Type Culture Collection (4 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Sk mel 2, by American Type Culture Collection (1 mentions)
Sk mel 1, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Plx4032, by Selleck Chemicals (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Actinomycin d, by Enzo Life Sciences (1 mentions)
Pd325901, by Selleck Chemicals (1 mentions)
Hek293, by Thermo Fisher Scientific (1 mentions)
Mccoy s 5a medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
U0126, by Merck Group (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Malme 3, by American Type Culture Collection (1 mentions)
Hek293t, by American Type Culture Collection (1 mentions)
Jurkat, by American Type Culture Collection (1 mentions)
6 protocols using sk mel 19
1
BRAF-Mutant Metastatic Melanoma Cell Lines
2
Metastatic Melanoma Cell Lines and Derivatives
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The metastatic melanoma cell lines A375, SK-MEL19 and SK-MEL28 were purchased from ATCC. SbCl2 cell line was a gift of Dr. B. Giovanelli (Stehlin Foundation for Cancer Research, St. Joseph Hospital, Houston, TX). The other metastatic melanoma cell lines used here and the vemurafenib resistant patient derived xenograft (PDX) cells, WM4205-3, were kindly gifted by M. Herlyn and C. Krepler from the Wistar Institute (Philadelphia, USA). These cells were tested every 6 month to exclude mycoplasm contaminations. The cell lines SbCl2 and SK-MEL2 carry an NRAS mutation, whereas all other cell lines used here are BRAF-mutated cells. All cell lines were cultured in RPMI 1640 medium with 10% fetal bovine serum (Sigma Aldrich) and 1% penicillin and streptomycin (Thermo Fisher Scientific). The MAPKi resistant melanoma cells were generated as previously described28 (link). The human primary melanocytes, fibroblasts and keratinocytes were isolated from human foreskin and cultured as previously described29 (link). Stat3 overexpressing cells and corresponding control cells of the SK-MEL19 cell line were generated as described before30 (link).
3
Culturing Melanoma Cell Lines
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4
Cytotoxicity Evaluation of Extracts
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The cytotoxicity of extracts was evaluated in vitro against one non-neoplastic cell line (MRC-5-human fibroblast) purchased from the American Type Culture Collection (ATCC) and two melanoma cell lines (SK-MEL-19 and SK-MEL-28) donated by Dr. María S Soengas (CNIO, Madrid, Spain). The cells were cultivated in 96-well plates (0.5 × 104 cells per well) and then were treated with extract in DMSO at a single concentration (50 μg/mL) over 72 h. The Alamar Blue™ assay was performed after 72 h following a standard procedure [53 (link)].
5
Cell Lines and Inhibitors for Melanoma Research
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The HEK293, Malme-3M, Malme-3 and A375 cell lines were obtained from American Type Culture Collection, and SK-Mel-19 was kindly provided by Dr. Neal Rosen (Memorial Sloan-Kettering Cancer Center). The cancer cell lines derived from melanoma (SK-Mel-19, A375 and Malme-3M) were maintained in RPMI-1640 medium (Gibco) supplemented with 10% heat-inactivated fetal bovine serum (Biological Industries), 50 U/ml penicillin, and 50 μg/ml streptomycin at 37°C with 5% CO2. The Malme-3 cells, which were derived from skin fibroblasts, were grown in McCoy's 5a Medium (Gibco) supplemented with 20% heat-inactivated FBS, 50 U/ml penicillin, and 50 μg/ml streptomycin at 37°C with 5% CO2. The human embryonic kidney cell line HEK293 was maintained in Dulbecco's modified Eagle's medium (Gibco) supplemented with 5% heat-inactivated fetal bovine serum, 50 U/ml penicillin, and 50 μg/ml streptomycin at 37°C with 5% CO2. The MEK inhibitors, PD325901, PLX4032 (Selleck) and U0126 (Sigma-Aldrich), and actinomycin D (Enzo) were dissolved in DMSO as stock solutions and stored at −20°C.
6
Authentication and Characterization of Cell Lines
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JURKAT (derived from T-ALL) and HEK 293T (derived from kidney) cell lines were purchased from the American Type Culture Collection (Manassas, VA). SUP-T1 (derived from T-LBL), PEER and HPB-ALL (derived from T-ALL) cell lines were obtained from the Leibniz Institute Deutsche Sammlung von Mikroorganismen und Zellkulturen-German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). SK-MEL-28 and SK-MEL-19 (derived from melanoma) (21) were obtained from Dr. Marisol Soengas (Centro Nacional de Investigaciones Ocológicas). The approximate doubling time of every cell line is 48 h, except for HEK-293T and SK-MEL-19 whose doubling time is 24 h. American Type Culture Collection, Deutsche Sammlung von Mikroorganismen und Zellkulturen and Centro Nacional de Investigaciones Ocológicas routinely perform cell line authentication using short tandem repeat profiling. Cell experimentation was always performed between 3 and 35 passages after thawing and cells were kept in culture not longer than 3 months. Mycoplasma tests were negative both at the beginning and at the end of experimentation.
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