Automated 3D Cell Culture and Screening by Imaging and Flow Cytometry

Three-dimensional (3D) cell cultures offer greater physiological relevance than monolayer cultures for cellular interaction studies and compound screens. However, manual manipulations of these cultures can be laborious and challenges are amplified as sample throughput increases. We used the Biomek FXP Workstation to automate the culture and drug sensitivity screening of cancer spheroids in Perfecta3D® Hanging Drop Plates. Automated steps include the plating of cells and addition of compounds to the hanging drops to induce apoptosis. Staining reagents were also added to analyze the spheroids by high content imaging. Finally, the transfer and dissociation of the spheroids into single cell suspensions was automated to enable flow cytometry analysis. Automated hanging drop cancer spheroids showed excellent consistency across wells and analyzing these spheroids by two complementary methods generated a more complete picture of drug responses in 3D cultures.