A375 melanoma cells

Manufactured by American Type Culture Collection

Sourced in United States

A375 melanoma cells are a commonly used in vitro model for studying melanoma, a type of skin cancer. These cells were derived from a human malignant melanoma and exhibit characteristics typical of melanoma cells. They can be used for various research applications involving melanoma biology and potential therapeutic interventions.

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A375 cells (54 citations) A375 melanoma (13 citations) A375 human melanoma cells (12 citations)

15 protocols using a375 melanoma cells

Culturing GFP-expressing Melanoma Cells

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A375 melanoma cells (ATCC) and GFP-expressing A375 human melanoma cells (Imanis Life Sciences; authenticated) were maintained in complete DMEM medium (ATCC) supplemented with 10% heat inactivated FBS (Gemini Bio-Products), 1% penicillin/streptomycin (ATCC), and 1 μg/mL puromycin to maintain high GFP expression (GIBCO).

Tyurina Y.Y., Kapralov A.A., Tyurin V.A., Shurin G., Amoscato A.A., Rajasundaram D., Tian H., Bunimovich Y.L., Nefedova Y., Herrick W.G., Parchment R.E., Doroshow J.H., Bayir H., Srivastava A.K, & Kagan V.E. (2023). Redox phospholipidomics discovers pro-ferroptotic death signals in A375 melanoma cells in vitro and in vivo. Redox Biology, 61, 102650.

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Cultivation of Melanocytes and Melanoma Cells

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Human melanocytes (ScienCell, Carlsbad, CA, HEM-l, Cat No 2200) were grown in MelM media containing MelGS growth supplements, 0.5% fetal bovine serum (FBS), and penicillin and streptomycin. A375 melanoma cells (ATCC CRL-1619) were grown in Complete Tu Medium containing a 4:1 mixture of MCDB-153 medium with 1.5 g/L sodium bicarbonate and Leibovitz’s L-15 medium with 2 mM L-glutamine, 2% FBS, and 1.68 mM CaCl₂. The cells were grown in a humidified tissue culture incubator at 37°C, 5% CO₂ atmosphere.

Zhao W., Mazar J., Lee B., Sawada J., Li J.L., Shelley J., Govindarajan S., Towler D., Mattick J.S., Komatsu M., Dinger M.E, & Perera R.J. (2016). The long non-coding RNA SPRIGHTLY regulates cell proliferation in primary human melanocytes. The Journal of investigative dermatology, 136(4), 819-828.

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Establishing GFP-Labeled Melanoma Cell Line

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A375 melanoma cells (ATCC) and RAW 264.7 macrophages (ATCC) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum and 1% penicillin/ streptomycin. Human dermal microvascular endothelial cells (HMVECs) and endothelial cell growth medium were from Promocell. Cells were incubated at 37 °C and in a humidified 5% CO₂ environment. The GFP-SR-B1 plasmid37 (link) was stably transfected in the A375 cells using Lipofectamine 2000 (Life Technologies) and transfectants were selected using Geneticin (Life Technologies) followed by fluorescent associated cell sorting (FACS).

Plebanek M.P., Mutharasan R.K., Volpert O., Matov A., Gatlin J.C, & Thaxton C.S. (2015). Nanoparticle Targeting and Cholesterol Flux Through Scavenger Receptor Type B-1 Inhibits Cellular Exosome Uptake. Scientific Reports, 5, 15724.

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Cell Culture Protocol for Leukemia and Melanoma

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All 293T cells, KG-1a myeloid leukemia cells, and A375 melanoma cells were purchased from the ATCC, and maintained in either DMEM or RPMI medium supplemented with 10% FBS, 1% L-Glutamine and 1% Penicilin/Streptamycin as described previously [33 (link)].

Kroeger C., Roesler R., Wiese S., Hainzl A, & Gatzka M.V. (2020). Interaction of Deubiquitinase 2A-DUB/MYSM1 with DNA Repair and Replication Factors. International Journal of Molecular Sciences, 21(11), 3762.

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Culturing eHAP and A375 Cells

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eHAP cells (Horizon Discovery Ltd, UK) were grown in IMDM supplemented with 10% FBS and 2 mM L-glutamine (all supplied by Gibco, UK). A375 melanoma cells (ATCC, USA) were thawed and cultured in DMEM supplemented with 10% FBS and 2 mM L-glutamine (all supplied by Gibco, UK). Cells were routinely checked for mycoplasma and identity verified by STR analysis.

Cross B.C., Lawo S., Archer C.R., Hunt J.R., Yarker J.L., Riccombeni A., Little A.S., McCarthy N.J, & Moore J.D. (2016). Increasing the performance of pooled CRISPR–Cas9 drop-out screening. Scientific Reports, 6, 31782.

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Culturing A375 Melanoma Cells

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2.1 Cell culture A375 melanoma cells (ATCC, Rockville, MD, USA) were cultured in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, Chicago, IL, USA), 100 U/ml penicillin/streptomycin (Gibco), and 4 mM L-glutamine (Corning, NY, USA) [36] (link).

Ercoli J., Finetti F., Woodby B., Belmonte G., Miracco C., Valacchi G, & Trabalzini L. (2020). KRIT1 as a possible new player in melanoma aggressiveness. Archives of biochemistry and biophysics, 691.

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A375 Melanoma Cell Culture

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A375 melanoma cells (ATCC, USA) were thawed and cultured in DMEM supplemented with 10% FBS and 2mM L-glutamine (all supplied by Gibco, UK). Cells were routinely checked for mycoplasma and identity verified by STR analysis.

le Sage C., Lawo S., Panicker P., Scales T.M., Rahman S.A., Little A.S., McCarthy N.J., Moore J.D, & Cross B.C. (2017). Dual direction CRISPR transcriptional regulation screening uncovers gene networks driving drug resistance. Scientific Reports, 7, 17693.

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Cell Culture Protocol for Cancer Cell Lines

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A375 melanoma cells, PC‐3 prostate cancer cells, and the human breast adenocarcinoma cell line MCF‐7 were all purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in Dulbecco's Modified Eagle's Medium with 4500 mg/L glucose, GlutaMAX–I, and pyruvate supplemented with 100 IU·mL⁻¹ penicillin/streptomycin and 10% fetal bovine serum. Cell media, cell culture consumables, antibiotics, trypsin‐EDTA, fetal bovine serum and PBS were purchased from Thermo Fisher Scientific Inc. (Rockford, IL, USA) unless otherwise stated.

Wallin H., Hunaiti S, & Abrahamson M. (2021). Externally added cystatin C reduces growth of A375 melanoma cells by increasing cell cycle time. FEBS Open Bio, 11(6), 1645-1658.

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Mangiferin's Anticancer Effects Evaluation

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Mangiferin was purchased from Shanghai Pureone Biological Technology Co., Ltd. (Shanghai, China). The purity of mangiferin was >95%, and the chemical structure is presented in Fig. 1A. The A375 melanoma cells, HepG2 and 7721 hepatocellular carcinoma cells, MCF-7 breast carcinoma cells, non-small cell lung cancer A549 cells, glioblastoma U87 cells and human embryonic lung fibroblast (HELF) cells were purchased from American Type Culture Collection (Manassas, VA, USA), and the normal human embryonic lung fibroblast (HELF) cell line was purchased from The Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Fetal bovine serum (FBS) was purchased from Gibco (Thermo Fisher Scientific, Inc., Waltham, MA, USA). 3-(4,5-dimetrylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), z-VAD-fmk (pan-caspase inhibitor), z-DEVD-fmk (caspase-3 inhibitor), z-LEHD-fmk (caspase-9 inhibitor), propidium iodide (PI) and rhodamine-123 were purchased from Sigma-Aldrich (St. Louis, MO, USA).

SHI W., DENG J., TONG R., YANG Y., HE X., LV J., WANG H., DENG S., QI P., ZHANG D, & WANG Y. (2016). Molecular mechanisms underlying mangiferin-induced apoptosis and cell cycle arrest in A549 human lung carcinoma cells. Molecular Medicine Reports, 13(4), 3423-3432.

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Establishment of Murine Tumor Models

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CHO cells, WEHI-164 fibrosarcoma cells, CTLL2 cells, F9 teratocarcinoma cells, CT26 colon carcinoma cells, L-M fibroblasts, HT1080 fibrosarcoma cells, A375 melanoma cells and C1498 acute myeloid leukemia chloroma cells were obtained from the American Type Culture Collection (ATTC) between 2015 and 2017, expanded and stored as cryopreserved aliquots in liquid nitrogen. Cells were grown according to the supplier’s protocol and kept in culture for no longer than 14 passages. Authentication of the cell lines also including check of post-freeze viability, growth properties and morphology, test for mycoplasma contamination, isoenzyme assay and sterility test were performed by the cell bank before shipment.
Eight week old female 129/SvEv mice, Balb/c mice and C57BL/6 were obtained from Janvier. Tumor cells were implanted subcutaneously in the flank using 15 x 10⁶ cells (F9), 5 x 10⁶ cells (WEHI-164), 2 x 10⁶ cells (CT26) and 1 x 10⁶ cells (C1498). Experiments were performed under a project license granted by the Veterinäramt des Kantons Zürich, Switzerland (27/2015).

De Luca R., Soltermann A., Pretto F., Pemberton-Ross C., Pellegrini G., Wulhfard S, & Neri D. (2017). Potency-matched dual cytokine-antibody fusion proteins for cancer therapy. Molecular cancer therapeutics, 16(11), 2442-2451.

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