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. 2011 Jun 23;2(6):e174. doi: 10.1038/cddis.2011.58

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Copyright © 2011 Macmillan Publishers Limited

This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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Spheroids are ‘primed for death'. BH3-profiling was performed on cells disaggregated from monolayers and spheroids grown from M28, REN, SARC and VAMT cells. The cells were gently permeabilized as described³⁹ and exposed to BH3 peptides (100 μM) for 90 min before JC1 was added for 30 min and depolarization measured as a percentage of the total. Cells from spheroids and monolayers were equally depolarized by Bim^BH3, the positive control, confirming a functioning apoptotic apparatus. However, cells disaggregated from spheroids were more sensitive to Bad^BH3, Hrk ^BH3 and ABT-737 than cells from monolayers, confirming that cells in 3D acquired apoptotic priming. Cells showed no response to an ABT-737 inactive enantiomer, used as negative control. (^*P<0.05 compared with monolayers, n=3)