This is a cell line derived from the murine acute myeloid leukemia C1498 cell line (ATCC® TIB-49^TM). Parental C1498 cells were transduced with LV-eGFP-PGK-Puro (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. A high eGFP expressing population was generated by selection using puromycin followed by selection using a methylcellulose based semi-solid medium.

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This cell line has been tested for mycoplasma contamination and is certified mycoplasma free.

The parental C1498 cell line was authenticated and certified free of interspecies cross contamination by STR profiling.

Due to the immunogenicity of the reporter genes in this cell line, we recommend using immunocompromised mice for in vivo studies.

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

Morphology

Cell photos taken at 200x magnification.

eGFP Expression

C1498-eGFP-Puro (green) or isotype control (C1498-Fluc-Puro; grey) cells were fixed with paraformaldehyde and analyzed by flow cytometry.

Complete Growth Medium: High glucose DMEM supplemented with 10% fetal bovine serum (FBS) and 1% Penicillin/Streptomycin.

To help maintain high eGFP expression, the cells can be subcultured in the presence of 1 μg/mL puromycin. It is also recommended that a backup frozen cell stock be generated before adding puromycin to the growth medium. Puromycin should not be added to the medium until a culture has been well established from the thawed cells (about 1 week).

The cells should be subcultured as needed to maintain a density between 3 x 10⁵ and 2 x 10⁶ cells/mL. The cells can be passaged by dilution in fresh complete growth medium (without centrifugation). However, regular passage using centrifugation as described above is recommended to limit the amount of debris in cultures.

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

C1498 cells form tumors and metastases at multiple sites, including the liver, lymph nodes, spleen, bone marrow, intestines, kidneys, and central nervous system, upon implantation into syngeneic C57BL/6 mice ^1,2.

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.

The cells can be amplified in vitro and used to generate additional frozen stocks. Cryopreservation of low passage stocks is recommended. Frozen stocks should be preserved in a designated cryopreservation medium.

These cells were generated via lentiviral vector transduction. The lentiviral vector used for transduction was a self-inactivating (SIN) vector in which the viral enhancer and promoter 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 LTR². Nevertheless, all work with these cells should be performed under biosafety-level 2 (BSL2) conditions by trained personnel. Institutional requirements may permit handling of these cells under BSL1 conditions if certain criteria are met.

References:

¹Boyer et al. Blood 1995 85: 2498-2506.

²Sauer et al. Cancer Res 2004 164 3914-3921.