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“Liver-macrophages, or Kupffer cells, are the resident macrophages this website in the liver [1]. These cells comprise approximately 10–15% of all liver cells [2]. These cells are localized along the sinusoidal space or its immediate vicinity in the liver, and are involved in the regulation of physiological homeostasis and immunosurveillance in this organ [3]. The liver-macrophages also play a protective role against liver damage and promote the regeneration

and fibrosis in cholestatic liver injury [1]. However, in pathological circumstances, activated liver-macrophages may aggravate liver damage, leading to cirrhosis and eventually failure of the organ. Therefore, liver-macrophages are considered to be an important

therapeutic target for pharmacological intervention [4,5]. Thus, various methods of isolating the liver-macrophages for in vitro studies have been elaborated and refined in a number of mammalian species. The conventional method of liver-macrophages starts with dissociation by collagenase perfusion, pronase treatment to remove parenchymal hepatocytes and finally counterflow centrifugal Ceritinib cost elutriation to separate the liver-macrophages from other nonparenchymal cells [6,7]. In addition, various modifications of this method for specific animal species have been reported, including human [8,9] and bovine [10,11] applications. However, all of these methods still require complex equipment and technical skill. To circumvent these technical difficulties, we recently developed a novel procedure for obtaining liver macrophages in sufficient number and purity, using a mixed primary culture of liver cells from the adult rat [12,13] and bovine [14]. In this study, we applied this simple and efficient Anidulafungin (LY303366) method to the neonatal swine liver and

succeeded repeatedly isolating sufficient numbers of swine liver-macrophages. These cells are useful tools for the functional analyses of the innate immune response in this important livestock species. Swine neonates at 1–7 days of age were obtained from the animal facility in the National Institute of Animal Health, according to the institutional guidelines for animal experiments (Approval no. 12-085). After deep anesthesia by intravenous injection of sodium pentobarbital (20 mg/kg) and exsanguination, the lobes of the swine liver were dissected out and parenchymal hepatocytes were isolated by the perfusion of saline followed by collagenase into the portal vein [10,15,16].