Fig. 1: Establishment of floating chicken enteroids.
From: Inside-out chicken enteroids with leukocyte component as a model to study host–pathogen interactions
a Matrigel-embedded embryonic chicken spheroids increase in size but lack budding at day 1 and b day 7 of culture. c Floating large multi-lobulated chicken enteroid structures develop over 3 days and d are maintained at day 9 of culture. Scale bar: 50 µm. e Time-lapse images from Supplementary video 1 showing the formation of budding crypt-like structures (marked by star *) in floating embryonic enteroids. Images are representative of data from at least 20 independent cultures each containing 2–3 embryos. Scale bar: 100 µm. Confocal images of floating embryonic enteroids showing proliferating cells (green, EdU) in both buds and core at f 14 h, g, h 3 days and i 7 days of culture. Images are representative of 3 independent cultures. Scale bar: 200 µm. j Quantification of floating embryonic enteroid numbers and morphology showed a significant increase in multiple budding enteroid numbers in the first day of culture. Data averaged for 3 cultures containing 2–3 embryos, 3 wells/culture, ~180 villi seeded per well at day 0. Villi: *t = 3.58, F = 8.07, df = 1; >1 bud enteroid: *t = 4.26, F = 0.03, df = 1 using a linear mixed effects model. Villi 0 h – 1 day: *p = 0.009, t = 10.65; >1 bud enteroid 0 h – 1 day: **p = 0.003, t = −18.92; >1 bud enteroid 5–7 day: *p = 0.008, t = 10.8 using post hoc pairwise t-tests.
