Species: Mouse

Cell type: Lymphoma

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

Media: RPMI, 10% FBS, 1% Pen/Strep

Description:

This is a cell line derived from the murine lymphoma A20 cell line (ATCC® TIB-208^TM). Parental A20 cells were transduced with LV-Fluc-P2A-Puro (Imanis #LV012) encoding the firefly luciferase (Fluc) cDNA under the spleen focus-forming virus (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 followed by selection using a methylcellulose based semi-solid medium. The lentiviral vector is 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 LTR¹.

Cell Line Authentication:

The parental A20 cell line was authenticated and certified free of interspecies cross contamination by STR profiling.

Recommended uses:

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

A20 cells form tumors and metastases at multiple sites, including the liver, lymph nodes, spleen, bone marrow, ovaries, and peritoneal cavity, upon implantation into syngeneic Balb/c mice⁶.

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.  

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

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.