The mini-guts in a culture well
To allow study of the relevance of for instance Paneth cell function and antimicrobial peptide secretion, as well as epithelial responses to a changed probiotic or prebiotic influence and the metabolic consequences thereof. The Paneth cells amount up to 10% of the epithelial culture-all other relevant specialized epithelial cell types are represented such as neuroendocrine cells. Clearly, these crypt cell based organoid cultures have as advantage that all relevant epithelial cell types (i.e. Paneth cells) are fully represented in the culture. Our group at AMC, in collaboration with the Gastroenterology Department, is further exploring the possibility to adapt the human crypt cell cultures as described earlier at different levels and there is a clear opportunity for Gut Research partnering to test for instance probiotic products in this system.
This culture setup allows us to study the direct effects of pre- and probiotics but also microbial metabolic factors that regulate epithelial and Paneth cell peptide expression and secretion of antimicrobial peptides in an easily manipulated ex vivo setting. Further, we can study the effect of inflammatory cytokines that intervene in transcriptional regulation of antimicrobial peptide expression and secretion. Lentiviral shRNA strategies are in place to study specific gene regulation of epithelial responses.
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