Culture media and format alter cellular composition and barrier integrity of porcine colonoid-derived monolayers
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Date
2024-04-02
Open Access Location
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Publisher
Taylor and Francis Group
Rights
(c) 2023 The Author/s
CC BY-NC-ND 4.0
CC BY-NC-ND 4.0
Abstract
Intestinal organoid technology has revolutionized our approach to in vitro cell culture due in part to their three-dimensional structures being more like the native tissue from which they were derived with respect to cellular composition and architecture. For this reason, organoids are becoming the new gold standard for undertaking intestinal epithelial cell research. Unfortunately, their otherwise advantageous three-dimensional geometry prevents easy access to the apical epithelium, which is a major limitation when studying interactions between dietary or microbial components and host tissues. To overcome this problem, we developed porcine colonoid-derived monolayers cultured on both permeable Transwell inserts and tissue culture treated polystyrene plates. We found that seeding density and culture format altered the expression of genes encoding markers of specific cell types (stem cells, colonocytes, goblets, and enteroendocrine cells), and barrier maturation (tight junctions). Additionally, we found that changes to the formulation of the culture medium altered the cellular composition of colonoids and of monolayers derived from them, resulting in cultures with an increasingly differentiated phenotype that was similar to that of their tissue of origin.
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Keywords
Barrier integrity, Porcine colonoids, colonoid-derived monolayers, differentiation, medium formulation, stem cells, Animals, Swine, Culture Media, Colon, Organoids, Intestinal Mucosa, Cell Culture Techniques
Citation
Barnett AM, Mullaney JA, McNabb WC, Roy NC. (2024). Culture media and format alter cellular composition and barrier integrity of porcine colonoid-derived monolayers.. Tissue Barriers. 12. 2. (pp. 2222632-).