Mel624

Manufactured by American Type Culture Collection

Sourced in United States

The Mel624 is a laboratory instrument for cell culture applications. It is designed to provide a controlled environment for the growth and maintenance of cells in vitro. The Mel624 offers precise temperature, humidity, and gas (CO2) regulation to support optimal cell culture conditions.

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Lab products found in correlation

9 protocols using mel624

Adoptive Transfer of Pmel-1 and OT-II T Cells

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Pmel-1 (B6.Cg-/Cy Tg [TcraTcrb] 8Rest/J), Rag2^−/−, OT-II (B6.Cg-Tg (TcraTcrb)425Cbn/J), and C57BL/6 mice were obtained from the Jackson Laboratory. C57BL/6 male mice of 6-8 weeks of age were used as recipient hosts for adoptive transfer unless otherwise indicated. We crossed Pmel-1 with Ly5.1 mice (B6.SJL-Ptprc^aPepc^b/BoyJ) to obtain Pmel-1 Ly5.1 mice. We crossed OT-II with Rag2^-/- to obtain OT-II Rag^-/- mice. All mice were maintained under specific pathogen–free conditions. B16 (H-2D^b), a murine melanoma, transduced as previously described31 (link) to express gp100 with human residues at position 25-27; EGS -> KVP. Mel624 was obtained from ATCC and Platinum-E ecotropic packaging cells were obtained from Cell Biolabs. Cell lines and maintained in DMEM media with 10% FBS, 1% glutamine and 1% penicillin-streptomycin.

Eil R., Vodnala S.K., Clever D., Klebanoff C.A., Sukumar M., Pan J.H., Palmer D.C., Gros A., Yamamoto T.N., Patel S.J., Guittard G.C., Yu Z., Carbonaro V., Okkenhaug K., Schrump D.S., Linehan W.M., Roychoudhuri R, & Restifo N.P. (2016). Ionic immune suppression within the tumour microenvironment limits T cell effector function. Nature, 537(7621), 539-543.

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Adoptive Transfer of Pmel-1 and OT-II T Cells

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Recombinant Protein Expression Strategies

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cDNAs encoding proteins in this study were purchased from Addgene or synthesized by Integrated DNA Technologies. All mutations were generated by PCR and tagged with either FLAG or HA or His, and cloned into vectors by Gibson assembly. pCDNA3.1(+) vector (Invitrogen) was used for transient expression; viral vectors (Clontech) for stable expression; and pET-28a (Novagen) for bacterial expression.
Wild-type, BRAF^−/− and CRAF^−/− MEFs were generated in previous study [58 (link), 59 (link)]. Melanoma cell lines: MeWo, A101D, Mel-624 were obtained from ATCC.

Yuan J., Ng W.H., Lam P.Y., Wang Y., Xia H., Yap J., Guan S.P., Lee A.S., Wang M., Baccarini M, & Hu J. (2018). The dimer-dependent catalytic activity of RAF family kinases is revealed through characterizing their oncogenic mutants. Oncogene, 37(43), 5719-5734.

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Human Cell Lines Cultivation Protocol

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Human cell lines 293 (Q-Biogene, Montreal, Canada), 293T (ATCC), 633 [55] (link), A375M [56] (link), C8161 [57] , Capan-1 (ATCC), HepG2 (ATCC), HMEC-1 (Life Technologies, Darmstadt, Germany), Mel624 [58] (link), Mel888 [58] (link), MiaPaCa-2 (ATCC), PANC-1 (ATCC), HUVEC (PromoCell, Heidelberg, Germany) and low passage human melanoma cells purified from human melanoma metastasis biopsies, PMelL, were cultivated using standard cell culture conditions as described before [14] (link).

Behr M., Kaufmann J.K., Ketzer P., Engelhardt S., Mück-Häusl M., Okun P.M., Petersen G., Neipel F., Hassel J.C., Ehrhardt A., Enk A.H, & Nettelbeck D.M. (2014). Adenoviruses Using the Cancer Marker EphA2 as a Receptor In Vitro and In Vivo by Genetic Ligand Insertion into Different Capsid Scaffolds. PLoS ONE, 9(4), e95723.

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Establishment of Fto-deficient MEF Cells

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Wild-type MEF (mouse embryonic fibroblasts) and Fto^−/− MEF cell lines were obtained from Prof. Arne Klungland. Spontaneously immortalized clonal MEF cell lines were established from individual E13.5-E15.5 days-old embryos obtained from heterozygous Fto^+/− crossings. Limbs were removed and remaining tissue chopped into small pieces and homogenized into single-cell suspension using a pipette. MEFs are grown in DMEM with 10% serum, 2 mM glutamine and penicillin/streptomycin. Cells grown for 4–7 days were trypsinized and frozen in complete medium with 10% DMSO as stock. Cells are grown in complete medium for 2–4 days before using them for following analysis. Human HeLa, HepG2, HEK293T, Mel624, and 3T3-L1 cell lines were purchased from ATCC. The aforementioned cells were maintained in DMEM (Gibco, #11965) with 10% fetal bovine serum (FBS, Gibco, 10438–026) and 1× Pen/Strep (Gibco, 15140) at 37°C with 5% CO₂. FTO stable knockdown and the knockdown control AML cells were obtained from Prof. Jianjun Chen and cultured as previously reported (Li et al., 2017 (link)).

Wei J., Liu F., Lu Z., Fei Q., Ai Y., He P.C., Shi H., Cui X., Su R., Klungland A., Jia G., Chen J, & He C. (2018). Differential m6A, m6Am, and m1A Demethylation Mediated by FTO in Cell Nucleus and Cytoplasm. Molecular cell, 71(6), 973-985.e5.

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Culturing Human Cell Lines

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Human melanoma cells A375, Mel-624, Mel-888, Human embryonic kidney cells HEK-293 and Human bladder epithelium immortalized cells SV-HUC-1 were all purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in complete DMEM containing 10% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA), 100 units of penicillin and 100 µg of streptomycin, at 37 °C in 5% CO2. Drug monensin sodium salt was purchased from Solarbio (Beijing, China) and dissolved in ethanol. All the procedures were in strict accordance with the Institutional Review Board of The Third Military Medical University.

Xin H., Li J., Zhang H., Li Y., Zeng S., Wang Z., Zhang Z, & Deng F. (2019). Monensin may inhibit melanoma by regulating the selection between differentiation and stemness of melanoma stem cells. PeerJ, 7, e7354.

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Culturing Melanoma Cell Lines

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The melanoma cell lines A375 and MEL624 were purchased from American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco’s modified Eagle's medium (DMEM) (Invitrogen Life Technologies, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Biomeda, Foster City, CA, USA) and 1% penicillin/streptomycin/glutamine (Gibco/Invitrogen, Carlsbad, CA, USA). All cell lines were maintained in a humidified incubator with 5% CO₂ at 37°C.

Yu Y., Xiang N., Lin M., Huang J.W., Zhang J., Cheng B, & Ji C. (2019). miR- 26a Sensitizes Melanoma Cells To Dabrafenib Via Targeting HMGB1-Dependent Autophagy Pathways. Drug Design, Development and Therapy, 13, 3717-3726.

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Culturing Melanoma Cell Lines

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The melanoma cell lines A375 and MEL624 were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (Biomeda Corp., Foster City, CA, USA) and 1% penicillin/streptomycin/glutamine (Thermo Fisher Scientific, Waltham, MA, USA). Cells were incubated in a humidified incubator with 5% CO₂ and 95% air at 37°C.

Ji C., Zhang Z., Chen L., Zhou K., Li D., Wang P., Huang S., Gong T, & Cheng B. (2016). Endoplasmic reticulum stress-induced autophagy determines the susceptibility of melanoma cells to dabrafenib. Drug Design, Development and Therapy, 10, 2491-2498.

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Culturing and Conditioning Human Melanoma Cells

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Human melanoma cells MeWo and FM3 were kindly provided by the Netherlands Cancer Institute (NKI), Mel603 by Leiden University Medical Center (LUMC). BLM cells were obtained from AIMM Therapeutics (Amsterdam, the Netherlands). Human melanoma cells A375, Mel-624, and the human lung epithelial carcinoma cell line A549 were authenticated by American Type Culture Collection (ATCC). All cell lines were tested mycoplasma free. Cell lines were maintained by culture in Dulbecco’s Modified Eagle Medium (DMEM, GIBCO) supplemented with 10% heat inactivated fetal bovine serum (FBS) and Antibiotic-Antimycotic (Thermofisher) in humidified incubators at 37°C and 5% CO₂. To obtain tumor-conditioned medium (CM), supernatant was collected after a four-day culture period or, when indicated, after a 24 h culture in serum-free medium. Conditioned medium was centrifuged at 1500 rpm for 5 min to pellet remaining cells and cell debris, aliquoted and stored at −20°C until further use.

Becker A.M., Decker A.H., Flórez-Grau G., Bakdash G., Röring R.J., Stelloo S., Vermeulen M., Piet B., Aarntzen E.H., Verdoes M, & de Vries I.J. (2024). Inhibition of CSF-1R and IL-6R prevents conversion of cDC2s into immune incompetent tumor-induced DC3s boosting DC-driven therapy potential. Cell Reports Medicine, 5(2), 101386.

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