Porcine hepatocyte–Kupffer cell co‐culture as an in vitro model for testing the efficacy of anti‐inflammatory substances
Hepatic cell culture models are considered as relevant and practical in vitro systems for studying the inflammatory response of the organ. Isolated and cultured hepatic cells contain all metabolizing enzymes and transport proteins, mimicking physiological and pathological processes that can occur in the liver in vivo (Herédi‐Szabó et al., 2009). Notwithstanding that primary cultures of hepatocytes also contain a distinct amount of non‐parenchymal cells, hepatocytes should be co‐cultured with KCs in appropriate ratio for proper investigations on hepatic inflammation. This functional cell culture system has been already established with cells of rat (Bhatia et al., 1998) and pig origin (Hoebe et al., 2000), serving as a model for human diseases and as a research tool in veterinary medicine. In the latter case, hepatocytes were co‐cultured with KCs in 1:1 ratio to study the hepatotoxicity of certain compounds (Hoebe et al., 2000).
The main goal of the present study was to improve and characterize a porcine primary hepatocyte–KC co‐culture to serve as an in vitro inflammatory model by applying different ratios of cell types to mimic milder and severe inflammation. These cell cultures might be useful to assess some aspects of enteral bacterial infections in swine as investigating the role of KCs in different forms of LPS‐induced inflammation. Further, the efficacy of potential anti‐inflammatory substances, such as the short‐chain fatty acid butyrate and terpinen‐4‐ol, the active component of the tea tree oil (derived from Melaleuca alternifolia) and further plants (such as Illicium verum and Myristica fragrans), was aimed to be tested on the co‐cultures. As both butyrate and terpinen‐4‐ol can be considered as potent natural anti‐inflammatory agents based on numerous in vitro and in vivo studies (Carson et al., 2006; Vieira, 2012; Farkas et al., 2014), they can be proper candidates to demonstrate new application possibilities of the obtained in vitro model.