In vitro models
Chicken 3D intestinal organoids
The chicken 3D intestinal organoids are based on the method described by Nash et al. (2021, 2023). The organoids are grown from intestinal villi isolated from 18-day-old embryos. The floating organoids are cultured in the absence of an extracellular matrix such as Matrigel, which leads to the formation of a model with multiple villus-like budding structures and an inside-out orientation. The externally accessible epithelial surface of the floating organoids allows for the uncomplicated testing of compounds and/or microbes. The intestinal organoids contain representative gut epithelial populations of stem cells, Paneth cells, enteroendocrine cells, goblet cells, and enterocytes.
Apart from the intact barrier function, the inner core of the 3D organoids is representative of the lamina propria and contains all cells that are present underneath the epithelial barrier, including immune cells such as natural killer cells, T cells and macrophages. The immune-epithelial component triggers appropriate immune responses making the organoids a suitable model for in vitro study of innate immune responses to enteric microbes and compounds.
These application make the 3D intestinal organoids an attractive model to test:
- food additives
- enzymes
- vaccines
- therapeutics
- genetics.
(Illustration: doi: 10.1038/s42003-021-01901-z)
Chicken 2D intestinal organoids
The chicken 2D intestinal organoid model is based on the method described in Orr et al. (2021). Similar to the 3D model, this method uses villi from 18-day-old embryos. The single cells are seeded on either plates or transwell inserts depending on the application. Similar to the 3D model, the 2D organoids contain representative gut epithelial populations of stem cells, Paneth cells, enteroendocrine cells, goblet cells, and enterocytes making this model more representative than a cell line.
The 2D organoid system allows for measurement of translocation of molecules including feed compounds across the epithelial layer. Furthermore, the integrity of the intestinal barrier can effectively be monitored by measuring Trans Epithelial Electrical Resistance (TEER) of FITC-dextran leakage in a time and dose dependent manner.
The 2D intestinal organoids can be used for the following applications:
- transwell barrier integrity studies
- intestinal absorption studies
- inflammatory response studies
- translocation studies (either passive or active transport)
- infection studies
(Illustration: doi: 10.1186/s13567-021-01010-z)
Our expert
Prof. dr. ir. Lonneke Vervelde
Senior scientist Poultry Health and R&D
Read the latest paper by Lonneke Vervelde about a new Enterococcus cecorum infection model at GD using chicken intestinal organoids: Invasion of Chicken Intestinal Cells Is Higher for Enterococcus cecorum Lesion Strains Compared to Cloacal Strains in an Organoid Model. Microorganisms2025, 13, 50