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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Multiplex quantitative analysis of stroma-mediated cancer cell invasion, matrix remodeling, and drug response in a 3D co-culture model of pancreatic tumor spheroids and stellate cells

Fig. 4

The effect of PSC co-culture on cell invasion from PANC-1 TSs. a Changes in size and shape of the spheroids under PSC co-culture conditions. b Three distinct stages of progressive invasion of cancer cells out of TSs. The cells with nuclear elongation are indicated by white arrowheads and nuclear aspect ratio was compared between cells remaining within spheroids and cells invading matrix. c Increased fraction of TSs with invasive morphology under PSC co-culture conditions. Invadopodia (yellow arrowheads) and dispersed single cells (yellow asterisks) can be seen around some spheroids. Sectional areas containing minimum of 30 spheroids were analyzed for their invasion stage of non-invasive, invadopodia, or invasive as defined in (b). The cell invasion was compared calculated the number of dispersed single cells found in the matrix outside spheroids and compared between mono-culture TSs and TSs co-cultured with PSCs on day 9 of culture. Nuclear DAPI (blue) or F-actin (red) staining was performed on cryo-sections of TS (a) or on whole TSs (b and c). Confocal optical sections were acquired at 1 μm (a and b) or 7 μm (c) intervals and stacked into a z-projection. TS: tumor spheroid. Data represent the mean ± SD of three independent experiments. Scale bars: 50 μm. ***p < 0.001

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