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Characteristic and Functional Analysis of a Newly Established Porcine Small Intestinal Epithelial Cell Line
- Source :
- PLoS ONE, PLoS ONE, Vol 9, Iss 10, p e110916 (2014)
- Publication Year :
- 2014
- Publisher :
- Public Library of Science (PLoS), 2014.
-
Abstract
- The mucosal surface of intestine is continuously exposed to both potential pathogens and beneficial commensal microorganisms. Recent findings suggest that intestinal epithelial cells, which once considered as a simple physical barrier, are a crucial cell lineage necessary for maintaining intestinal immune homeostasis. Therefore, establishing a stable and reliable intestinal epithelial cell line for future research on the mucosal immune system is necessary. In the present study, we established a porcine intestinal epithelial cell line (ZYM-SIEC02) by introducing the human telomerase reverse transcriptase (hTERT) gene into small intestinal epithelial cells derived from a neonatal, unsuckled piglet. Morphological analysis revealed a homogeneous cobblestone-like morphology of the epithelial cell sheets. Ultrastructural indicated the presence of microvilli, tight junctions, and a glandular configuration typical of the small intestine. Furthermore, ZYM-SIEC02 cells expressed epithelial cell-specific markers including cytokeratin 18, pan-cytokeratin, sucrase-isomaltase, E-cadherin and ZO-1. Immortalized ZYM-SIEC02 cells remained diploid and were not transformed. In addition, we also examined the host cell response to Salmonella and LPS and verified the enhanced expression of mRNAs encoding IL-8 and TNF-α by infection with Salmonella enterica serovars Typhimurium (S. Typhimurium). Results showed that IL-8 protein expression were upregulated following Salmonella invasion. TLR4, TLR6 and IL-6 mRNA expression were upregulated following stimulation with LPS, ZYM-SIEC02 cells were hyporeponsive to LPS with respect to IL-8 mRNA expression and secretion. TNFα mRNA levels were significantly decreased after LPS stimulation and TNF-α secretion were not detected challenged with S. Typhimurium neither nor LPS. Taken together, these findings demonstrate that ZYM-SIEC02 cells retained the morphological and functional characteristics typical of primary swine intestinal epithelial cells and thus provide a relevant in vitro model system for future studies on porcine small intestinal pathogen-host cell interactions.
- Subjects :
- Lipopolysaccharides
Salmonella typhimurium
Carcinogenesis
Cellular differentiation
Veterinary Microbiology
Sus scrofa
Cell
Cell Culture Techniques
lcsh:Medicine
Apoptosis
Epithelium
Intestinal mucosa
Molecular Cell Biology
Intestinal Mucosa
lcsh:Science
Telomerase
Mice, Inbred BALB C
Multidisciplinary
Cell biology
Jejunum
medicine.anatomical_structure
Veterinary Diseases
Host-Pathogen Interactions
Cytokines
Anatomy
Cellular Types
Genetic Engineering
Research Article
Biotechnology
Histology
Immunology
Mice, Nude
Bioengineering
Biology
Microbiology
Cell Line
Immune system
Cell Line, Tumor
medicine
Animals
Humans
Secretion
Cell Shape
Cell Proliferation
Tissue Engineering
Cell growth
lcsh:R
Biology and Life Sciences
Epithelial Cells
Cell Biology
Molecular Development
Small intestine
Biological Tissue
Cell culture
lcsh:Q
Veterinary Science
Biomarkers
Neoplasm Transplantation
Developmental Biology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 9
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....b4aec7e8ccaaae10f93589979859d253