Species: Mouse

Cell type: Lewis Lung Carcinoma

Transgenes: Murine sodium iodide symporter (mNIS) 

Media: DMEM, 10% FBS, 1% Pen/Strep, 2μg/mL puromycin

Description: LL/2-mNIS-Puro is a monoclonal population of the Lewis lung carcinoma cell line LL/2 (ATCC® CRL-1642™), also commonly known as LLC1, transduced with LV-mNIS (LV008) encoding the murine sodium iodide symporter (mNIS) cDNA under the spleen focus-forming virus (SFFV) promoter. The clones were selected by limiting dilution after transduction of parental LL/2 cells with LV-mNIS. Clones were expanded and mNIS function confirmed using a I-125 uptake assay.

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: The parental LL/2 cell line was authenticated and certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 27 STR loci.

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 in your lentiviral transduced cells.

In vivo: LL/2 cells form tumors post implantation into immunocompromised or syngeneic C57BL/6 mice. The in vivo growth of these metastases can be monitored using noninvasive, high-resolution 3D PET/SPECT imaging.

 

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

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.

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.