Stable Transfection of Murine NIS in B16F10 Melanoma and DHD/K12/TRb Colorectal Cancer Cells

The B16F10 melanoma cells (obtained from ATCC) and the DHD/K12/TRb colorectal cancer cells (obtained from Sigma) were cultured in DMEM (GIBCO, France) supplemented with 10% heat-inactivated fetal calf serum (FCS, GIBCO) at 37°C under a humidified atmosphere containing 5% CO₂. Cells were passaged by using 0.05% trypsin. Both cell lines were transfected with pcDNA3.1-mNIS (murine NIS)36 (link) using the FuGENE 6 reagent (Roche) according to the manufacturer’s instructions. Stable clones were selected by addition of 1 mg/mL geneticin (G418) to the medium 3 days after transfection. One clone with high functional expression of NIS was selected for each cell line.37 (link)

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Le Goas M., Paquet M., Paquirissamy A., Guglielmi J., Compin C., Thariat J., Vassaux G., Geertsen V., Humbert O., Renault J.P., Carrot G., Pourcher T, & Cambien B. (2019). Improving 131I Radioiodine Therapy By Hybrid Polymer-Grafted Gold Nanoparticles. International Journal of Nanomedicine, 14, 7933-7946.

Publication 2019

B16F10 melanoma cells DMEM FCS

Atmosphere Cell line Cells Clone Colorectal cancer Fugene 6 G418 Geneticin Melanoma Murine Transfection Trypsin

Corresponding Organization : Institut de Biosciences et Biotechnologies

Other organizations : Commissariat à l'Énergie Atomique et aux Énergies Alternatives, Université Paris-Saclay, Centre Antoine Lacassagne

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Variable analysis

independent variables

Transfection of B16F10 melanoma cells and DHD/K12/TRb colorectal cancer cells with pcDNA3.1-mNIS (murine NIS)

dependent variables

Functional expression of NIS

control variables

Culture medium: DMEM supplemented with 10% heat-inactivated fetal calf serum
Cell culture conditions: 37°C, humidified atmosphere with 5% CO2
Cell passaging method: 0.05% trypsin

positive controls

None specified

negative controls

None specified

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