Species: Mouse

Cell type: C57/L hepatoma

Transgenes: Firefly luciferase (Fluc) and neomycin resistance (Neo) for selection with G418

Media: DMEM, 10% FBS, 1% Pen/Strep, 1.25 mg/mL G418

Description:

Hepa1-6-Fluc-Neo is a polyclonal population derived from the hepatoma Hepa1-6 cell line (ATCC® CRL-1830^TM) transduced with a lentiviral vector LV-Fluc-P2A-Neo (LV011) encoding the firefly luciferase (Fluc) cDNA under the spleen focus-forming virus (SFFV) promoter linked to the neomycin resistance gene (Neo) 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:

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 luciferase expressing cell line suitable for use as a positive control cell line in bioluminescence assays to verify luciferase 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. The Fluc transgene facilitates in vivo noninvasive bioluminescent imaging of implanted cells. Fluc is immunogenic and may cause tumor rejection in immunocompetent mice. For the most consistent results, immunocompromised mice are recommended for studies.

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

Luciferase Assay: 10⁴, 10⁵, or 10⁶ cells were placed in wells of a 96-well plate and 0.3 mg of d-luciferin was added. The plate was immediately imaged using a Xenogen IVIS Spectrum.