Immunochemical, biomolecular and biochemical characterization of bovine epithelial intestinal primoculturesReport as inadecuate




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BMC Cell Biology

, 6:42

First Online: 01 December 2005Received: 24 August 2005Accepted: 01 December 2005

Abstract

BackgroundCultures of enterocytes and colonocytes represent valuable tools to study growth and differentiation of epithelial cells. In vitro models may be used to evaluate passage or toxicity of drugs, interactions of enteropathogenes bacteria strains with intestinal epithelium and other physiologic or pathologic phenomenon involving the digestive tract.

ResultsCultures of bovine colonocytes and jejunocytes were obtained from organoid-enriched preparations, using a combination of enzymatic and mechanical disruption of the intestine epithelium, followed by an isopicnic centrifugation discarding most single cells.

Confluent cell monolayers arising from plated organoids exhibited epithelium typical features, such as the pavement-like structure, the presence of apical microvilli and tight junctions. Accordingly, cells expressed several markers of enterocyte brush border i.e. maltase, alkaline phosphatase and fatty acid binding protein as well as an epithelial cytoskeleton component cytokeratin 18. However, enterocyte primocultures were also positive for the vimentin immunostaining mesenchyme marker. Vimentin expression studies showed that this gene is constitutively expressed in bovine enterocytes. Comparison of the vimentin expression profile with the pattern of brush border enzymes activities, suggested that the decrease of cell differentiation level observed during the enterocyte isolation procedure and early passages of the primoculture could result from a post-transcriptional de-repression of vimentin synthesis. The low differentiation level of bovine enterocytes in vitro could partly be counteracted adding butyrate 1–2 mM or using a glucose-deprived culture medium.

ConclusionThe present study describes several complementary approaches to characterize bovine primary cultures of intestinal cells. Cultured cells kept their morphologic and functional characteristics during several generations.

Electronic supplementary materialThe online version of this article doi:10.1186-1471-2121-6-42 contains supplementary material, which is available to authorized users.

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Author: Dorina Rusu - Suzanne Loret - Olivier Peulen - Jacques Mainil - Guy Dandrifosse

Source: https://link.springer.com/article/10.1186/1471-2121-6-42



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