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HCT116-hNIS-Neo/eGFP-Puro

Species: Human

Cell type: Colorectal carcinoma

Transgenes: Human sodium iodide symporter (hNIS) with neomycin resistance (Neo) for selection with G418 and enhanced green fluorescent protein (eGFP) with puromycin resistance for selection with puromycin

Media: DMEM, 10% FBS, 1% Pen/Strep, 0.5 mg/mL G418, 1μg/mL puromycin

Description: HCT116-hNIS-Neo/eGFP-Puro is a polyclonal population of the human colorectal carcinoma cell line HCT116 (ATCC® CCL-247™) transduced with 1) LV-hNIS-Neo (LV013) encoding the human sodium iodide symporter (hNIS) cDNA under the spleen focus-forming virus (SFFV) promoter linked to the neomycin resistance gene (Neo) via an IRES and 2) LV-eGFP-PGK-Puro (LV031) encoding enhanced green fluorescent protein (eGFP) cDNA under the SFFV promoter and the puromycin resistance gene (Puro) under the PGK 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).

Mycoplasma Testing: This cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

Cell Line Authentication: Authentication of the parental HCT116 cell line was performed by short tandem repeat (STR) profiling with 9 STR loci including CSF1PO, D13S317, D16S539, D5S818, D7S820, TH01, TPOX, vWA and sex chromosome marker Amelogenin. STR profiling of HCT116 cells are verified and there is no interspecies cross contamination detected.

Recommended uses:

In vitro: This is a high hNIS/eGFP expressing clone suitable for use as a positive control cell line in iodine uptake assays and fluorescence assays to validate NIS or GFP expression respectively in your lentiviral transduced cells.

In vivo: This human colorectal cell line grows as xenografts in immunocompromised mice. Growth of these tumors can be monitored using noninvasive high-resolution 3D SPECT or PET imaging or optical imaging for GFP fluorescence.

Note: In life optical imaging GFP fluorescence is not recommended due to high background autofluorescence. Tissues may be harvested post mortem for analysis by conventional microscopy. 

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)

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

 

Flow Cytometry for GFP: HCT116-hNIS-Neo/eGFP-Puro (green) and control (gray) 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

Why Choose Imanis?

With quality reagents, services, and support, we have you covered at all stages of your study. Our expert scientists can help you select the best reagents for your needs and offer tips to optimize your experiments and achieve superior results. Our wide-range of reporter gene and oncolytic virus reagents undergo rigorous quality control testing so you can be confident in the quality of the products you receive. And with our competitive prices, you can complete your study for less.

  • “Our group has, and continues to, use NIS as a noninvasive reporter for cell transplantation studies in mice and in pigs. Imanis has provided expert technical and analytical support for this research, and has allowed us to publish our research in high impact journals, including Science Translational Medicine.” – Dr. Raymond Hickey, Mayo Clinic

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Use Fewer Animals

Our Reduction Campaign

At Imanis, we are committed to promoting the practice of the 3Rs in animal research. Learn how we are decreasing the use of animals and research as well as saving up to 15% on your orders. Continue reading...