List of Cell Lines
|Cell specification||Entity of origin|
Assay procedure. Tumor cells autonomously assemble to form spheroids in round-bottom 96 well plates. Compounds incubate for three days before readout of cell quantity with either cell viability reagent CellTiter-Glo-3D or luciferase measurement.
Example of U87MG mono-spheroids. Left: Comparison of the size of U87MG spheroids at day 0 and 3. Right: Concentration-response curve of trametinib treatment on U87MG mono-spheroids. Positive control (0 % viability) is staurosporine treatment; negative control (100 % viability) is vehicle control. ‘day 0’ shows the number of viable cells at the start of treatment.
Assay procedure for co-culture spheroids: Firefly-luciferase expressing tumor cells and renilla-expressing stroma cells are seeded in a round-bottom 96 well plate and left to assemble for spheroid formation for three days. Compounds incubate for three days before firefly-luciferase and renilla-luciferase quantification from the lysate of the spheroid. The cytotoxic and anti-proliferative effects of compounds can be determined for both tumor and stroma cells separately.
Example showing reduced sensitivity of DLD1 cells against trametinib in co-spheroids: DLD1 tumor cells were cultured in spheroids as mono-culture (black) or together with HS27A stroma cells (blue). Spheroids were treated for 72 h with the MEK kinase inhibitor trametinib. Subsequently, the viability of tumor cells was analyzed by measurement of firefly luminescence.
- Co-spheroids with tumor & stroma cells, luciferase measurement only in tumor cells
- Co-spheroids with firefly luciferase-labeled tumor & renilla-luciferase labeled stroma cells
Setup: IC50 value determination and single concentration testing are possible.
Custom-tailored assays can be performed.
Cell lines that are not part of the list can be established according to the client’s research needs.
Controls: DMSO only treatment serves as high control. Staurosporine treatment (at 1x10-5M) serves as low control.
Turnaround time: Results will be available after 3 to 4 weeks. Expedited scheduling and data delivery can be arranged prior to study commencement.
Report: A detailed report including assay conditions, methodology, and comprehensive evaluation of data as well as raw data for each analysis will be provided. Photographs are optional.
Screening Facility: The assay is performed in Freiburg, Germany.
Compound requirements: In brief: 30 µl of the stock solution with 1000x of the highest assay concentration or the equivalent of solid material.