Human melanoma cells SK-MEL-28, SK-MEL-2, and SK-MEL-5 (ATCC) and mouse cell line CT26 (ATCC) were cultured in monolayers using RPMI supplemented with 10% heat inactivated bovine serum (Thermo Fisher Scientific), 10mM L-glutamine (Corning), and 0.5% penicillin G-streptomycin sulfate (Corning). Cells were detached using 0.25% trypsin EDTA (Corning) for passaging. The murine melanoma cell line D4M3A was generated from Tyr::CreER;BrafCA;Ptenlox/lox mice (13 (link)) and kindly provided by Dr. David Mullins (Dartmouth University, Hanover, NH). D4M3A cells were cultured as previously described (13 (link)). All cells were low-passage and confirmed to be mycoplasma-free (LookOut mycoplasma kit; Sigma).
Penicillin g streptomycin sulfate
Manufactured by Corning
Penicillin G-streptomycin sulfate is a combined antibiotic solution commonly used in cell culture media. It provides broad-spectrum antimicrobial protection to inhibit the growth of a variety of bacteria and fungi.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Corning (3 mentions)
L glutamine, by Corning (2 mentions)
Heat inactivated bovine serum, by Thermo Fisher Scientific (2 mentions)
Lookout mycoplasma kit, by Merck Group (2 mentions)
Sk mel 2, by American Type Culture Collection (1 mentions)
Sk mel 5, by American Type Culture Collection (1 mentions)
Sk mel 28, by American Type Culture Collection (1 mentions)
Mts cell proliferation colorimetric assay kit, by Abcam (1 mentions)
3 protocols using penicillin g streptomycin sulfate
1
Culturing Human and Murine Melanoma Cells
2
Melanoma Cell Culture and Viability Assay
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Human melanoma cell lines SK-MEL-2, SK-MEL-5, SK-MEL-28, M14, and LOX-IMVI were obtained from ATCC (Manassas, VA). Cells were cultured in monolayers using RPMI supplemented with 10% heat-inactivated bovine serum (Thermo Fisher Scientific), 10mM L-glutamine (Corning), and 0.5% penicillin G-streptomycin sulfate (Corning). Cells were detached using 0.25% trypsin EDTA (Corning) for passaging and were cultured at 37°C in 5% CO2. The murine melanoma cell line D4M3A was generated from Tyr::CreER; BrafCA; Ptenlox/lox mice22 and kindly provided by Dr. David Mullins (Dartmouth University, Hanover, NH). D4M3A cells were cultured as described in Jenkins et al., 2014. All cells used in experiments were low-passage and were confirmed to be mycoplasma-free (LookOut mycoplasma kit; Sigma). During cell viability assays, cells were plated in 96-well plates and treated with T-VEC after 12–16 hr. Cell viability was measured using MTS assay and cell viability measured according to the manufacturer’s instructions (MTS Cell Proliferation Colorimetric Assay Kit, Biovision, Milpitas, CA).
3
Cell Culture Protocols for Cancer Research
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The CT26 and B16F10 cell lines were obtained from Dr. Andrew Zloza (Rush University). The D4M3A cells were provided by Dr. David Mullins (Dartmouth University). Cells were cultured as monolayers at 37 degrees Celsius and 5% CO2 in RPMI (CT26 and D4M3A) or DMEM (B16F10) supplemented with 10% heat-inactivated bovine serum and 0.5% penicillin G-streptomycin sulfate (Corning). 0.25 trypsin EDTA (Corning) was used to detach adherent cells.
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