Organoids were propagated in hPDAC media modified from Boj et al.66 (link). For drug screening, organoids were dissociated to single cells and seeded on top of a thin layer of Matrigel (Corning, NY, USA) in 384-well plate (3000/well). Next day, organoids were treated with a range of MK-2206 (0.05–30 µM) and Trametinib (0.001 nM to 10 µM) or SHP099 (0.05–400 µM) concentrations for 96 h and cell viability was determined by CellTiter-Glo 3D viability assay (Promega, Madison, USA). Drug-response curves were graphed and half maximal inhibitory concentration values were calculated using GraphPad Prism 8.0 (San Diego, USA).
Gebregiworgis T., Kano Y., St-Germain J., Radulovich N., Udaskin M.L., Mentes A., Huang R., Poon B.P., He W., Valencia-Sama I., Robinson C.M., Huestis M., Miao J., Yeh J.J., Zhang Z.Y., Irwin M.S., Lee J.E., Tsao M.S., Raught B., Marshall C.B., Ohh M, & Ikura M. (2021). The Q61H mutation decouples KRAS from upstream regulation and renders cancer cells resistant to SHP2 inhibitors. Nature Communications, 12, 6274.
Other organizations :
Princess Margaret Cancer Centre, University Health Network, University of Toronto, Diamond Materials (United States), Purdue University West Lafayette, Center for Cancer Research, University of North Carolina at Chapel Hill
Cell viability determined by CellTiter-Glo 3D viability assay
control variables
Organoids seeded on top of a thin layer of Matrigel in 384-well plate (3000/well)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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