Establishment of immortalized rat Kupffer cell lines

Background. Kupffer cells have been implicated in the pathogenesis of various liver diseases. Primary cultures of Kupffer cells have a very limited life span, tend to de-differentiate and become senescent, and therefore are not suitable for cell signaling studies. Aim. To establish immortalized rat Kupffer cell lines that facilitate mechanistic studies of cell signaling and signal transduction. Methods. Rat Kupffer cells were sub-cultured with EGF to obtain rat Kupffer Cell line 1 (RKC1), and subsequently transfected with Simian Virus 40 Large T-antigen expression vector to obtain rat Kupffer Cell line 2 (RKC2). Results. RKC1 and RKC2 are similar to primary Kupffer cells as they express the molecular markers ED1, ED2, ED3, and F4/80, and upregulate TNF-α, IL-6, IL-1β, Fas /FasL, and NF-κB, as well as TLR4 in response to LPS or pancreatic elastase. Additionally, RKC1 and RKC2 maintain phagocytic properties of latex beads and exhibit increased telomerase and stabilized p53 activity. Conclusion. Immortalized RKC1 and RKC2 cells maintain properties of primary Kupffer cells and can be valuable tools in evaluating the role of Kupffer cells in immune diseases and in liver-cell based drug discovery.