title: The Ex Vivo Colon Organ Culture and Its Use in Antimicrobial Host Defense Studies.
abstract: The intestine displays an architecture of repetitive crypt structures consisting of different types of epithelial cells, lamina propia containing immune cells, and stroma. All of these heterogeneous cells contribute to intestinal homeostasis and participate in antimicrobial host defense. Therefore, identifying a surrogate model for studying immune response and antimicrobial activity of the intestine in an in vitro setting is extremely challenging. In vitro studies using immortalized intestinal epithelial cell lines or even primary crypt organoid culture do not represent the exact physiology of normal intestine and its microenvironment. Here, we discuss a method of culturing mouse colon tissue in a culture dish and how this ex vivo organ culture system can be implemented in studies related to antimicrobial host defense responses. In representative experiments, we showed that colons in organ culture express antimicrobial peptides in response to exogenous IL-1β and IL-18. Further, the antimicrobial effector molecules produced by the colon tissues in the organ culture efficiently kill Escherichia coli in vitro. This approach, therefore, can be utilized to dissect the role of pathogen- and danger-associated molecular patterns and their cellular receptors in regulating intestinal innate immune responses and antimicrobial host defense responses.
authors.affiliation: Department of Pathology, UT Southwestern Medical Center.
authors.forename: S M Nashir
authors.initials: SM
authors.lastname: Udden
authors.affiliation: Department of Pathology, UT Southwestern Medical Center.
authors.forename: Sumyya
authors.initials: S
authors.lastname: Waliullah
authors.affiliation: Department of Pathology, UT Southwestern Medical Center.
authors.forename: Melanie
authors.initials: M
authors.lastname: Harris
authors.affiliation: Department of Pathology, UT Southwestern Medical Center; Hasan.Zaki@utsouthwestern.edu.
authors.forename: Hasan
authors.initials: H
authors.lastname: Zaki
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000494:pharmacology*; D003106:Colon / Q000166:cytology* / Q000378:metabolism; D015169:Colony Count, Microbial; D003470:Culture Media / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects*; D007113:Immunity, Innate; D007413:Intestinal Mucosa / Q000166:cytology* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009924:Organ Culture Techniques
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D003470:Culture Media