This is a polyclonal population derived from the EMT6 mammary carcinoma cell line (ATCC® CRL-2755™). Parental EMT6 cells were transduced with LV-eGFP-PGK-Puro (Imanis #LV031), encoding the enhanced green fluorescent protein (eGFP) cDNA under the spleen focus-forming virus (SFFV) promoter and the puromycin resistance gene (Puro) under the phosphoglycerate kinase (PGK) promoter. High eGFP expressing cells were selected 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.

*The ATCC trademark and trade name and any and all ATCC catalog numbers are trademarks of the American Type Culture Collection.

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.

Replication Competent Lentivirus (RCL) Test (including a test report) is available for this cell line at an added cost.  Contact us to learn more.

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

In vivo Imaging

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A Nude mouse was implanted with 2.5 x 10⁵ EMT6-eGFP-Puro (CL153) cells in the right hind flank. Tumor growth was monitored over time using calipers. Fluorescence imaging was performed on Day 22 using an IVIS Spectrum.

Morphology

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Cell photos taken at 200x magnification.

Fluorescence Expression

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EMT6-eGFP-Puro (green) or isotype control (EMT6-Fluc-Puro; grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry.

Complete Growth Medium: 

Dulbecco’s Modifies Eagles Media (DMEM), 10% fetal bovine serum (FBS), 1% Penicillin/Streptomycin, 1 μg/mL puromycin (to maintain high eGFP 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.

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.