Species: Human

Cell type: Fibrosarcoma

Transgenes: Near-infrared fluorescent protein (iRFP; excitation/emission: 690/713nm) with puromycin resistance (Puro) for selection with puromycin

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

Description: HT1080-iRFP-Puro is a polyclonal population derived from the fibrosarcoma cell line HT1080 (ATCC® CCL-121™) transduced with a lentiviral vector 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 vectors used are self-inactivating (SIN) vectors 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: HT1080-iRFP-Puro cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

Cell Line Authentication: The parental HT1080 cell line was authenticated are certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 9 STR loci including CSF1PO, D13S317, D16S539, D5S818, D7S820, TH01, TPOX, vWA and sex chromosome marker Amelogenin.

Recommended uses:

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

In vivo: HT1080 cells form tumors post implantation into immunosuppressed mice. The in vivo growth of these tumors can be monitored using noninvasive optical imaging for iRFP.

Please ensure that your optical imaging system has the correct excitation and emission filters for detection of iRFP (excitation/emission: 690/713nm).

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

Flow Cytometry for iRFP: HT1080-iRFP-Puro (red) or control (HT1080-Fluc-Puro) cells (grey) were fixed with paraformaldehyde and analyzed by flow cytometry (20,000 events).

Recommendations for noninvasive in vivo imaging: use the NIS reporter gene for high resolution 3D SPECT or PET in vivo 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.