Cell Culture


hours prior to cell viability assays. For cell viability assays, cells were assayed in the spheroid microplate using CellTiter-Glo® 3D (Promega Cat. No. G9683) following vendor’s protocols. Luminescence was detected using a PerkinElmer EnVision™ Multilabel plate reader.


Dose response curves of chemotherapeutic com- pounds applied to mono- and co-culture spheroids for 48 hours were generated using CellTiter-Glo 3D cell viability assay (Figure 4). The data were normalised to spheroids in the presence of 0.5% DMSO as 100% viability control.


Toxicity potency (TC50) values of chemothera- peutic compounds applied to mono- and co-culture spheroids for 48 hours were generated using CellTiter-Glo 3D cell viability assay (Table 2). Although the presence of endothelial cells did not affect the potency of Paclitaxel on HT-29 colon tumour cells, a right-shift in the potency was


observed with Vinorelbine, and a left-shift in potency was observed with Cisplatin under co-cul- ture conditions.


Conclusion In vitro 3D cell culture models are now becoming increasingly complex, and their usefulness in sup- porting cell growth, tissue morphogenesis, stem cell differentiation, disease modelling, drug dis- covery and toxicity testing is growing. It is evi- dent that a range of 3D models with varying physiological characteristics are required to meet the needs of specific cell types or assays. Currently, many complex 3D models of tumorige- nesis, stem cell differentiation and organoid for- mation employ naturally-derived ECM-based hydrogels. Combining newer technologies, such as microarray on a chip, microfluidics or even bio-printing with biologically relevant materials,


Figure 3 Dose response curves of chemotherapeutic compounds applied to mono-, co- and tri- culture spheroids for 48 hours using CellTiter-Glo® 3D cell viability assay. The presence of fibroblast (FB) and immune cells affected the potency of several compounds. Black = A549, blue = FB, red = A549 + FB and green = A549 + FB + PBMC


Drug Discovery World Winter 2017/18


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