Excellent signal/background ratio and stable luciferase expression make this cell line ideal for in vivo bioluminescence imaging of xenograft animal model to study human cancer and monitor activity of anti-cancer drug. It also can be used in cell-based assays for cancer research.

The base medium for this cell line is ATCC-formulated RPMI-1640 Medium (ATCC 30-2001). To make the complete growth medium, add the following components to the base medium:

• Fetal bovine serum (FBS; ATCC 30-2020) to a final concentration of 10%
• Blasticidin to a final concentration of 8 µg/mL

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
   Cultures can be established between 2 x 10⁴ and 4 x 10⁴ viable cells/cm².
6. Incubate cultures at 37°C.

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