Abstract
Recently, 3D small intestinal organoids (enteroids) have been developed from cultures of intestinal stem cells which differentiate in vitro to generate all the differentiated epithelial cell types associated with the intestine and mimic the structural properties of the intestine observed in vivo. Small-molecule drug treatment can skew organoid epithelial cell differentiation toward particular lineages, and these skewed enteroids may provide useful tools to study specific epithelial cell populations, such as goblet and Paneth cells. However, the extent to which differentiated epithelial cell populations in these skewed enteroids represent their in vivo counterparts is not fully understood. This study utilises label-free quantitative proteomics to determine whether skewing murine enteroid cultures toward the goblet or Paneth cell lineages results in changes in abundance of proteins associated with these cell lineages in vivo. Here, proteomics data confirms that skewed enteroids recapitulate important features of the in vivo gut environment, demonstrating that they can serve as useful models for the investigation of normal and disease processes in the intestine. Furthermore, comparison of mass spectrometry data with histology data contained within the Human Protein Atlas identifies putative novel markers for goblet and Paneth cells.
Highlights
3D small intestinal organoids have been developed from enterocytes, goblet cells, enteroendocrine cells, M cells, and Paneth cells.[1,2] Gobcultures of intestinal stem cells which differentiate in vitro to generate all the let and Paneth cells play crucial roles differentiated epithelial cell types associated with the intestine and mimic the structural properties of the intestine observed in vivo
Comparison of mass spectrometry data with histology data contained within the Human Protein
The small intestine is organized into protruding finger-like villi, application of unbiased profiling techniques, such as proteomics, and crypts of Lieberkuhn which extend into the muscularis mu- to study these cells
Summary
This study utilises label-free quantitative proteomics to determine whether skewing murine enteroid cultures toward the goblet or Paneth cell lineages results in changes in abundance of proteins associated with these cell have been shown to play a role in luminal antigen sampling across the small intestinal epithelium.[5] Paneth cells reside at the crypt base, and secrete antimicrobial compounds into the crypt lulineages in vivo. Wastling Faculty of Natural Sciences Keele University cell fate, giving rise to differentiated epithelial lineages These renewal properties can be exploited to generate 3D organoid cultures (called enteroids) from isolated intestinal LGR5+ stem cells or crypts.[12] When cultured in Matrigel R with a cocktail of growth factors, LGR5+ stem cells generate the differentiated epithelial.
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