This is a polyclonal population derived from the EMT6 mammary carcinoma cell line (ATCC® CRL-2755™). Parental EMT6 cells were transduced with LV-Fluc-P2A-Puro (Imanis #LV012) encoding the firefly luciferase (Fluc) cDNA under the SFFV promoter and linked to the puromycin resistance gene (Puro) via a P2A cleavage peptide. A high Fluc expressing population was generated by selection using puromycin. The lentiviral vectors are self-inactivating (SIN) vectors in which the viral enhancers and promoters have 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 LTR¹.

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This cell line has been tested for mycoplasma and found negative for mycoplasma contamination.

The parental EMT6 cell line has been authenticated and certified free of interspecies cross contamination by short tandem repeat (STR) profiling.

¹Miyoshi et al. J Virol. 1998. 72:8150-8157.

In vivo Imaging

Blab/c mice were implanted intravenously with 5×10⁵ EMT6-Fluc-Puro (CL154) cells. Tumors were imaged by BLI on Day 10.

Morphology

Cell photo taken at 200x magnification.

Luciferase Expression:

The indicated number of cells were placed in wells of a 96-well plate. After the addition of 3 mg/mL d-luciferin, the plate was immediately imaged using an IVIS Spectrum. The total flux (photons/sec) was plotted as a function of cell number.

Complete Growth Medium: Dulbecco’s Modifies Eagles Media (DMEM), 10% fetal bovine serum (FBS), 1% Penicillin/Streptomycin, 1 μg/mL puromycin (to maintain high Fluc expression)

Typical commercial puromycin stocks are provided at a concentration of 10 mg/mL or 10,000X. Puromycin should NOT be added to the medium until a culture has been well established from the thawed cells (about 1 week). It is also recommended that a backup frozen cell stock be generated before adding puromycin to the growth medium.

These cells are suitable for in vitro and in vivo experimentation.

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.