Comprehensive Chemotherapeutic Compound Screening

A library of 627 commercially available chemotherapeutic and targeted oncology compounds were tested at 5 concentrations in 10-fold dilutions. The library consisted of 180 approved drugs, 334 investigational compounds and 113 probes (Supplementary Dataset 1). The chemical compounds, DMSO (negative control) and benzethonium chloride (positive controls) were added to 384-well plates using an acoustic liquid dispensing system ECHO 500/550 (Labcyte). Biobanked frozen MNCs were thawed, resuspended in CM (conditioned medium) constituted of 77.5% RPMI 1640, 10% FCS, 12.5% human HS-5 bone marrow stromal cell line derived conditioned medium and 1% penicillin and streptomycin, let recover for 3 h, and live cells counted. Compounds were first dissolved by adding 5 μL of cell free medium, followed by 20 μL cell suspension containing 5000 viable cells to each well using EL 406 plate washer-dispenser (BioTek). The plates were incubated at 37 °C in 5% CO₂ for 72 h. Subsequently, CellTiter-Glo (Promega) reagent was added to all wells and cell viability was measured using a PHERA star FS multimode plate reader (BMG Labtech).
The drug responses passing the data quality assessment were included in further analysis.^{70 (link)} Drug sensitivity scores (DSS) were calculated as shown previously.^{44 (link)}