This is a polyclonal population derived from the prostate PC3 cell line (ATCC® CRL-1435^TM). Parental PC3 cells were transduced with LV-Fluc-P2A-Puro (Imanis #LV012) encoding the firefly luciferase (Fluc) cDNA under the spleen focus-forming virus (SFFV) promoter linked to the puromycin resistance gene (Puro) via a P2A cleavage peptide. High Fluc expressing cells were selected using puromycin.

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This cell line has been tested negative for mycoplasma contamination.

The parental PC3 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.

In vivo Imaging

A Nude mouse was implanted with 2.5 x 10⁵ PC3-Fluc-Puro (CL122) cells in the right hind flank. Tumor growth was monitored over time using calipers. Bioluminescence imaging was performed on Day 70 using an IVIS Spectrum.

Morphology

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 a Xenogen 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, 3 mg/mL puromycin.

For maintenance, a subcultivation ratio of 1:3 is recommended. At this ratio cells will be ready for passage approximately every 3-4 days.

Cells can be amplified and used to generate additional frozen stocks. Frozen stocks should be preserved in a designated cryopreservation medium or in complete growth medium without puromycin supplemented with 5-10% DMSO.

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

The Fluc transgene facilitates in vivo noninvasive bioluminescent imaging of implanted cells.

These cell 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 LTR¹.  All work with these cells should be performed under biosafety-level 2 (BSL2) conditions by trained personnel.

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