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Fig. 2 | BMC Cancer

Fig. 2

From: Recapitulating epithelial tumor microenvironment in vitro using three dimensional tri-culture of human epithelial, endothelial, and mesenchymal cells

Fig. 2

a Confocal image of the synthetic tumor microenvironment mimic (STEM) spheroid after staining for live and dead cells (Green color represents calcein AM staining indicating live cells, and red represents ethidium homodimer staining indicating dead cells) (Scale bar – 200 μm). b (i) Immunostaining of STEM at the end of day 15 for hypoxia marker pimonidazole. Hypoxia was confirmed by antibody binding (pink color) which is prominent in the interior of the STEM. The nuclei were counter-stained with DAPI. (ii) Scoring of proliferation and hypoxia within various regions of the STEM. The scoring was adapted from Mikhail et al.[25]. c Fluorescent micrographs of STEMs generated using turbo GFP expressing human pulmonary microvascular endothelial cells (HPMECs), turbo RFP expressing A549, and MSCs, which turbo GFP and turbo RFP negative cells, i.e. only DAPI positive. Cell nuclei were stained blue using DAPI nuclear stain. DAPI positive, GFP negative and RFP negative cells in the merged image represent MSC populations (represented by blue color) (Scale bar 100 μm)

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