- Frozen (CL075) $ 1,250
Cell type: Fibrosarcoma
Transgenes: Enhanced green fluorescent protein (eGFP) with puromycin resistance (Puro) for selection with puromycin
Media: DMEM, 10% FBS, 1% Pen/Strep, 1 μg/mL puromycin
Description: HT1080-eGFP-Puro is a polyclonal population from the fibrosarcoma cell line HT1080 (ATCC® CCL-121™) transduced with a lentiviral vector LV-eGFP-PGK-Puro (LV031) encoding the enhanced green fluorescent protein (eGFP) cDNA under the spleen focus-forming virus (SFFV) promoter and the puromycin resistance gene (Puro) under the phosphoglycerate kinase promoter.
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-eGFP-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.
In vitro: This is a high eGFP expressing cell line suitable for use as a positive control cell line in fluorescence assays to verify eGFP expression in your lentiviral transduced cells.
In vivo: HT1080 cells form tumors post implantation into immunosuppressed mice. The presence of these tumors may be detected using optical imaging for eGFP at post mortem analysis. Please note that noninvasive eGFP imaging in living animals will be suboptimal due to high autofluorescence and poor light penetration.
For in vivo imaging, please 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.
Morphology: Low- and high-density cell morphology (200x)
Flow Cytometry for eGFP: HT1080-eGFP-Puro (green) or control (HT1080-Fluc-Puro) cells were fixed with paraformaldehyde and analyzed by flow cytometry (20,000 events).
Publications that used associated reagents:
LV-eGFP-PGK-Puro (LV031): Shen et al. Immunovirotherapy with vesicular stomatitis virus and PD-L1 blockade enhances therapeutic outcome in murine acute myeloid leukemia. Blood. 2016. March 17: 127(11): 1449-58.