Species: Human

Cell type: Lung Carcinoma

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

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

Description: A549-eGFP-Puro is a polyclonal population of the human lung carcinoma cell line A549 (ATCC® CCL-185™) transduced with LV-eGFP-PGK-Puro (LV031) encoding 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: The A549-eGFP-Puro cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

Cell Line Authentication: The parental A549 cell line was authenticated and certified free of interspecies cross-contamination by short tandem repeat (STR) profiling with 9 STR loci including CSF1PO, D13S317, D16S539, D5S818, D7S820, TH01, TPOX, vWA and sex chromosome marker Amelogenin.

Recommended uses:

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

In vivo: A549 cells form metastases in the lungs of mice post systemic administration. The in vivo growth of these metastases can be monitored using optical imaging.

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

Flow Cytometry for eGFP: A549-eGFP-Puro (green) or control (A549-Fluc-Puro; grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry (20,000 events).

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.