Journal: Communications Biology
Article Title: A simple high-throughput approach identifies actionable drug sensitivities in patient-derived tumor organoids
Figure Lengend Snippet: Mini-ring approach to unveil drug response patterns in PDTOs. a Morphology of all PDTOs established in this study as visualized by bright-field microscopy. Morphology and 3D organization of the samples is highly variable. For instance, some of Patient #3 cells are arranged in fascicles within the Matrigel, likely representing the sarcomatous component of the tumor. Scale bar, 100 µm. b Results of kinase screening experiment for Patient #1 PDTOs. Three readouts were used for this assay: ATP quantification as measured by CellTiter-Glo 3D and organoid number or size quantification evaluated by bright-field imaging. Bright-field images were segmented and quantified using the Celigo S Imaging Cell Cytometer Software. Both organoid number and total area were evaluated for their ability to capture response to drugs. In this plot, each vertical line is one drug, all 240 tested are shown. Values are normalized to the respective vehicle controls for each method and expressed as %. Average Z-score calculated as reported in Methods. c A representative image of the effects of the indicated drug treatments as visualized by the Celigo cell imager. Scale bar, 100 µm. d Small-scale kinase assay on Patient #1 primary PDTOs and PDX-derived cells. ATP readout. Four molecules not present in the primary screening were tested. Flavopiridol and BS-181 HCl are included as positive and negative control, respectively. t -test, ** p
Article Snippet: ATP assay After the organoid release with dispase as indicated above, 75 µl of CellTiter-Glo 3D Reagent (Promega #G968B) is added to each well followed by 1 min of vigorous shaking.
Techniques: Microscopy, Imaging, Cytometry, Software, Kinase Assay, Derivative Assay, Negative Control