Fig. 1

Generation and expression profiling of porcine PSCdMs. Schematic and timeline illustrating the differentiation protocol and cytokines used for deriving macrophages from porcine PSCs. Solid arrows indicate steps in which cells are attached: either on STO feeder cells (PSCs), gelatin (macrophage differentiation) or non-coated TC plastic (macrophage maturation). The hollow arrow representing mesoderm induction indicates that embryoid body formation was performed in suspension. Representative bright-field images are shown for the different cell morphologies generated at each stage. B RT-qPCR expression profile analysis of cells generated at each step of macrophage differentiation for markers of pluripotency (NANOG), early mesoderm induction (KDR1), HSC induction (RUNX1) and macrophages (PU.1 and CSF1R). Mean and SD of two biological replicates from three experiments. C Score plot showing the first two principal components (PC1 and PC2) in tissue-specific gene expression. Based on PC1 and PC2 the data shows good separation of porcine PSCs, in vitro-derived porcine PSCdMs, ex vivo-derived PAMs, microglia, brain and other tissue samples. D Heatmap of the hundred most highly expressed genes in pig tissues and cell lines. Lower expression levels are highlighted in blue and higher expression values in red. Biological replicates are indicated in green. Hierarchical clustering of the samples, shown as a tree at the top of the heatmap, was calculated using Euclidean distances between samples after transposing the variance stabilised expression data. E Heatmap showing the similarities between samples based on the Euclidean distances. The darker the colour, the closer the sample relationship is based on their expression profile. F Flow cytometry analysis comparing primary pig PAMs with in vitro-derived porcine macrophages derived from two independent porcine PSC lines (PSCdM 1 & 2) co-stained with CD14/CD16 (red) and CD169/CD172a (blue) relative to isotype controls (grey)