|ZHANG, Q - Purdue University|
|APPLEGATE, T - Purdue University|
|AJUWON, K - Purdue University|
Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 4/10/2015
Publication Date: 7/27/2015
Citation: Zhang, Q., Applegate, T.J., Eicher, S.D., Ajuwon, K. 2015. Validation of a chicken ileal explant culture for measurement of mucosal inflammation induced by lipopolysaccharide. Poultry Science Association Meeting Abstract. No. 342P.
Technical Abstract: Gut mucosa holds a single layer of epithelial cells and the largest mass of lymphoid tissue in the body. While epithelial cell cultures are widely used to assess intestinal barrier functions, they have limitations to study cellular interactions with other cells, in particular those of the immune system. In this study, a chicken ileal explant culture was validated for investigating short-term mucosal inflammation in an ex vivo environment. Initially, ileal explants from broilers at 21 d of age were cultured in vitro up to 6 h. As assessed by lactate dehydrogenase (LDH) released into the culture medium, explants within 2 h of incubation remained over 90% viable. Also, from the morphological scores, the explants cultured by 2 h were considered as an acceptable duration for short-term testing of inflammation. Subsequently, lipopolysaccharide (LPS) dose-related responses were determined for ileal explants cultured by 2 h. The results from LDH in culture medium showed that the viability of ileal explants was remarkably decreased when more than 50 µg/mL LPS were added (P < 0.05). A significant nitric oxide release was observed when 10 and 20 µg/mL LPS were applied (P < 0.05), respectively. Also, the highest inflammatory response was detected at 20 µg/mL LPS based on gene expressions of TLR-4 (LPS recognition), iNOS, TNF-a, IL-1ß, IL-8 (inflammatory cytokines), Claudin-1, Claudin-5 (tight junction proteins) as well as MUC2, IgA and PIgR (secretory immune system). These results demonstrate the potential usefulness of this intestinal explant culture for studying cellular interactions as well as evaluating nutrient biological effects in the state of inflammation, but only suitable for rapid responses.