Immunochemical, biomolecular and biochemical characterization of bovine epithelial intestinal primocultures

摘要: Cultures of enterocytes and colonocytes represent valuable tools to study growth differentiation epithelial cells. In vitro models may be used evaluate passage or toxicity drugs, interactions enteropathogenes bacteria strains with intestinal epithelium other physiologic pathologic phenomenon involving the digestive tract. bovine jejunocytes were obtained from organoid-enriched preparations, using a combination enzymatic mechanical disruption intestine epithelium, followed by an isopicnic centrifugation discarding most single Confluent cell monolayers arising plated organoids exhibited typical features, such as pavement-like structure, presence apical microvilli tight junctions. Accordingly, cells expressed several markers enterocyte brush border (i.e. maltase, alkaline phosphatase fatty acid binding protein) well cytoskeleton component (cytokeratin 18). However, primocultures also positive for vimentin immunostaining (mesenchyme marker). Vimentin expression studies showed that this gene is constitutively in enterocytes. Comparison profile pattern enzymes activities, suggested decrease level observed during isolation procedure early passages primoculture could result post-transcriptional de-repression synthesis. The low partly counteracted adding butyrate (1–2 mM) glucose-deprived culture medium. present describes complementary approaches characterize primary cultures Cultured kept their morphologic functional characteristics generations.