Species: Mouse

Cell type: Lewis Lung Carcinoma

Transgenes: Near infrared fluorescent protein (iRFP; ex/em = 690/713 nm) with puromycin resistance (Puro) for selection with puromycin.

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

Description: LL/2-iRF-Puro is a polyclonal population of the Lewis lung carcinoma cell line LL/2 (ATCC® CRL-1642™), also commonly known as LLC1, transduced with LV-iRFP-P2A-Puro (LV032) encoding the near-infrared fluorescent protein (iRFP) cDNA under the spleen focus-forming virus (SFFV) promoter linked to the puromycin resistance gene (Puro) via a P2A cleavage peptide.

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).

Mycoplasma Testing: The LL/2-iRFP-Puro cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

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 iRFP expressing cell line suitable for use as a positive control cell line in a fluorescence assay to verify iRFP expression in your lentiviral transduced cells.

In vivo: LL/2 cells form tumors post implantation into immunosuppressed mice. The in vivo growth of these tumors can be monitored using noninvasive optical imaging.

For in vivo imaging, please use the nonimmunogenic murine NIS reporter gene.

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)

Flow Cytometry for iRFP: LL/2-iRFP-Puro (red) or isotype control (LL/2; grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry (20,000 events).