Species: Mouse

Cell type: C57/L hepatoma

Transgenes: Enhanced green fluorescent protein (eGFP) and puromycin resistance (Puro) for selection with puromycin

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

Description: Hepa1-6-eGFP-Puro is a polyclonal population derived from the hepatoma Hepa 1-6 cell line (ATCC® CRL-1830^TM) transduced with 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 under control of 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: The parental Hepa1-6 cell line was authenticated and cells are certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 27 STR loci. 

Recommended uses:

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

In vivo: Hepa1-6 cells form primary tumors and spontaneous lung metastases post implantation into syngenic C57L/J mice.

Due to high background autofluorescence in animals, eGFP is not recommended for whole animal in-live imaging. Rather, samples can be collected post mortem for analysis by conventional fluorescence microscopy or flow cytometry. eGFP is immunogenic and may cause tumor rejection in immunocompetent mice. For the most consistent results, immunocompromised mice are recommended for studies. 

For whole animal in vivo imaging, please use luciferase, iRFP, or the nonimmunogenic murine NIS reporter gene.

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

Transgene Validation: Flow cytometry of Hepa1-6-eGFP-Puro (green) and isotype control (Hepa1-6; gray) cells. 

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.