Species: Mouse

Cell type: Melanoma

Transgenes: Firefly luciferase (Fluc) with puromycin resistance (Puro) for selection with puromycin

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

Description: B16F10-Fluc-Puro is a polyclonal population of the murine melanoma cell line B16F10 (ATCC^® CRL-6475^™) transduced with LV-Fluc-P2A-Puro (LV012) encoding the firefly luciferase (Fluc) cDNA under the spleen focus-forming virus (SFFV) promoter linked to the puromycin resistance gene (Puro) via a P2A cleavage peptide.

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).

Cell Line Authentication: The parental B16F10 cell line was authenticated and 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 Fluc 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: B16F10 cells can form subcutaneous tumors after flank implantation or metastases in the lungs of mice after intravenous administration. The in vivo growth of these metastases can be monitored using bioluminescent imaging with D-luciferin substrate.

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 to the indicated wells. The plate was immediately imaged using a Xenogen IVIS Spectrum.

 

Bioluminescent images showing growth of a subcutaneous B16-Fluc tumor in an athymic mouse. 

 

 

 

 

 

 

 

 

10⁷ B16-Fluc-Puro cells were injected subcutaneously into the right flanks of female NCR athymic mice. Mice were imaged at day 0 on the day of cell implantation and at day 7 using a Perkin Elmer IVIS® Spectrum system, at 10-15 minutes post intraperitoneal injection of D-luciferin at 150 mg/kg. Tumor size was measured using calipers. Data from a representative mouse (ID#6383) is shown.