Fig. 6

Morphological characterisation of sequential seeded co-cultured spheroids. OvCar8 and Detroit 551, A2780 and Detroit 551 co-cultured spheroids were seeded time-shifted and cultured for 96 h as described in Fig. 2A. (A,B) Spheroids were examined by scanning electron microscopy after 96 h of growth. Scale bar 50 μm. (C,D) Prior to cultivation, the cells were stained with different fluorescence dyes in order to subsequently assign them to a cell type during cultivation. These spheroids were fixed, cleared and imaged after 96 h of growth using LSM. Red/Cell Tracker Deep Red: ovarian cancer cells; green/CellTracker Green CMFDA: fibroblasts. Scale bar 200 μm. (E,F) Histological and immunohistochemical analysis of spheroid morphology and protein expression of spheroids after 96 h of growth. Ki-67 is a proliferation marker, while PAX8 is marker for tumour cells and fibroblast activation protein markers fibroblasts, respectively. Scale bar 200 μm.