Isolation and characterization of human primary enterocytes from small intestine using a novel method.

摘要: Cell culture studies of enterocytes are important in many fields. However, there difficulties obtaining cell lines from adult human intestine, such as microbial contamination cultures the tissue samples, short life span enterocytes, overgrowth mesenchymal cells, etc. Various model used to obtain intestinal very complex requiring use feeder layer or gel matrices. The aim this study was establish a novel method for simple and reproducible isolation enterocytes. Enterocytes were isolated SI samples (n = 5) obtained cadaveric donors using mechanical procedure, separation with immunomagnetic beads coated anti-EpCAM antibodies. Light electron microscopy, flow cytometry immunocytochemistry techniques characterize cells. Immunohistochemical staining normal SB biopsies confirmed that maintained an vivo phenotype reflected cytokeratin expression CK18, CK20 intestine-specific markers sucrase isomaltase maltase glucoamylase. Furthermore, cells strongly expressed TLR-5, 6, 7, 8 10 several molecules CD40, CD86, CD44, ICAM-1 HLA-DR which triggering cell-mediated immune responses. This technique provides unique vitro system biology conditions well inflammatory processes various small bowel disorders.