Figure 2. Low-throughput and high-throughput drug testing in cancer organoid lines and treatment efficacy determination, and summary of human primary liver cancer specimens, demographics, and laboratory derivation of sister PDO lines.

(A) A panel of 7 cancer drugs at a concentration of 10 μM, along with negative control (0.1% DMSO) and a positive control (10% Triton X-100), were used on a PDO line. The PDO treated with cisplatin appeared identical to the negative control, consistent with no drug efficacy. Gemcitabine stalls PDO growth, but does not kill cells. We verified that gemcitabine does not induce cell death through additional studies, as described in Supplemental Figure 2. (B) Cisplatin allows unrestricted growth of cancer PDOs — as demonstrated by continuously expanding cystic PDO structures — while gemcitabine had a cytostatic effect. The efficacy of bortezomib, a proteasome inhibitor, was validated in a time-course experiment. The cystic PDO structures were effectively prevented from maintaining their shape or expanding in size. (C) The most frequently used clinical drug combinations for CCA were used on a CCA PDO line. As shown, gemcitabine plus cisplatin had the same effect as gemcitabine alone. Similarly, in all 8 combinations tested, the overall efficacy was similar to that of the more efficacious of the 2 drugs used in combination. Scale bars: 200 μm (A) and 400 μm (B and C).