Incucyte live cell technology

All fresh-collected specimens were obtained after informed consent of parents and patients and were anonymized for their analyses. This study was conducted in accordance to the local ethical committee approval (declaration number: DC-2017-3090). All patients and their paired-lines and xenografts are detailed in Table 1. As depicted in Figure 1 To prepare the 2D and 3D experiments, doubling time calculations were systematically done in an incubator (HERAcell VIOS 250i, CO2 incubator, Thermofischer), where oxygen concentrations were modulated from 1 to 21%. The cell growth was then followed for 7 days using IncuCyte ® Live Cell technology (Essen BioScience). Pictures were automatically acquired over time and quantified with IncuCyte ® software. Cell population doubling time was determined using the following equation: doubling time (days)=ln2(t-t0)/ln(Nt/N0), where t-t0 is the time of exponential growth, Nt and N0 are cell numbers at time t and t0, respectively.
For all experiments in single culture or in co-culture (see section 4.6), real-time scanned contrastphase images were acquired and analyzed by IncuCyte ® ZOOM Live Cell Analysis system (Essen BioScience).