Species: Mouse

Cell type: Melanoma

Transgenes: Murine sodium iodide symporter (mNIS) 

Media: DMEM, 10% FBS, 1% Pen/Strep

Description: B16F10-mNIS-Puro is a monoclonal population of the murine melanoma cell line B16F10 (ATCC^® CRL-6475^™) transduced with LV-mNIS (LV008) encoding the murine sodium iodide symporter (mNIS) cDNA under the SFFV promoter.

The lentiviral vector used is a self-inactivating (SIN) vector in which the viral enhancer and promoter has been deleted. Transcription inactivation of the LTR in the SIN provirus increases biosafety by preventing mobilization by replication competent viruses and enables regulated expression of the genes from the internal promoters without cis-acting effects of the LTR (Miyoshi et al., J Virol. 1998).

Cell Line Authentication: Authentication of the parental B16F10 cell line was performed by short tandem repeat (STR) profiling with 27 STR loci. STR profiling of B16F10 cells are verified and there is no interspecies cross contamination detected.

Recommended uses:

In vitro: This is a high mNIS expressing cell line suitable for use as a positive control cell line in iodine uptake assays to verify NIS expression in your lentiviral transduced cells.

In vivo: B16F10 cells form metastases in the lungs of mice. The in vivo growth of these metastases can be monitored using noninvasive, high-resolution 3D PET or SPECT imaging.

References on NIS imaging:

1. Fruthwirth et al. A whole body dual modality radionuclide optical strategy for preclinical imaging of metastasis and heterogeneous treatment responses in different. microenvironments. J. Nucl. Med 2014. 55(4): 686-94.

2. Penheiter et al. The sodium iodide symporter (NIS) as an imaging reporter for gene, viral and cell-based therapies. Curr Gene Ther. 2012, 12(1):33-47.

Morphology: Low- and high-density cell morphology (200x)

Transgene Validation:

NIS Function Assay (Iodine Uptake): Cells were incubated with ¹²⁵I for 1h in the presence or absence of KClO₄, an inhibitor of iodine uptake. Radioiodine concentrated within the cells was measured with a gamma counter.