Figure 5.

Infiltration-kinetics of PBMC toward rSFV-infected cancer-spheroids

(A) The setup of a cancer-spheroid and PBMC co-culture experiment to quantify immune infiltration in a 3D model using microscopy. (B) Representative microscopy images of far-red stained PBMC (in red) associated with LNCaP (top row) or PANC-1 (bottom row) spheroids at day 4 post co-culture. Kinetics of PBMC infiltration in (C) LNCaP or (D) PANC-1 spheroids infected with rSFV-particles up to 4 days post co-culture. See Figure S5 to visualize the microscopy images of different replicates for respective conditions. As a positive control, T cells stimulated overnight with antibodies against CD3 and CD28 were used. In (B and C) the plots represent data from 8 replicate-spheroids and 2 independent donors. The number of far-red positive PBMC is quantified as the number of red events per image, where each image consists of an individual spheroid per well. Data are presented as mean ± SEM.