Fig. 6: Composition and functionality of immune cells within chicken enteroids.
From: Inside-out chicken enteroids with leukocyte component as a model to study host–pathogen interactions
a–i Confocal images of chicken enteroids stained for leukocyte markers (green) at 2 or 7 days of culture. All enteroids are counterstained with DAPI (blue) and Phalloidin (red). a–c Enteroids stained for CD45 (green) showing leukocytes in the a, c lamina propria and b epithelium. Enteroids at 2 days of culture stained for d CD3, e CD4, f CD8β, g chB6, h TCR2 (chicken αβ1 TCR), i and TCR3 (chicken αβ2 TCR). j–k Enteroids cultured from CSF1R-eGFP transgenic embryos at j day 2 and (k1 magnification of k) day 7 of culture. Scale bar: 20 µm. l Expression of immune cell-related genes in freshly isolated villi (0 h), 3 day and 7 day chicken enteroids was compared by RNA sequencing analysis. Heat maps show the expression levels (log2 counts per million reads) of a range of immune cell-related genes. RNA sequencing data is representative of 3 independent experiments, each comprising of 2 technical replicates, each containing 3 embryos. m–p pHrodo zymosan bioparticles (red) were added to 3 day enteroid cultures and visualised m in the epithelium after 8 h and n in CD45+ leukocytes (green) after 48 h. (o1 magnification of o) Zymosan was also added to 7 day enteroid cultures and phagocytosis of zymosan by leukocytes in the ‘lamina propria’ core was visualised after 48 h. p Zymosan uptake was blocked in 3 day enteroid cultures treated with Cytochalasin D, which inhibits actin polymerization. Scale bar: 20 µm. Images are representative of data from at least 3 independent cultures each containing 2–3 embryos.
