Cell-based Assay

1. Process: firstly, the sensitivity screening of single drug is carried out, and then cell lines that are moderately sensitive or resistant to the two compounds are selected for the drug combination study; the two compounds can be used for the study through fixed dose or fixed ratio.

2. Cells: resuscitate the cells and incubate them for one week before use; continuous incubation should not exceed 4 weeks;

3. Inoculation time: adherent cells need to be inoculated one day before dosing, while suspension cells can be inoculated on the day of dosing or one day in advance;

4. Inoculation density: for 72hr treatment, typically inoculate 3000-6000 cells/well/100 uL of medium; if longer treatment is required, the inoculation density needs to be reduced or optimized;

5. Processing time: generally 72hr, or two cell doubling times, or the time can be specified by clients;

6. Compound concentration: set the concentration range based on the IC50 value determined during individual administration;

7. Repetition: no need for repetition; can be repeated in whole microplate;

8. Detection reagent: Promega CellTiter-Glo® (CTG) assay

9. Data results: single drug IC50, Combination Index (CI)