Cocultures Kupffer cell-hepatocyte

Tukov FF, Maddox JF, Amacher DE, Bobrowski WF, Roth RA, Ganey PE. Modeling inflammation-drug interactions in vitro A rat Kupffer cell-hepatocyte coculture system. Toxicol In Vitro. 2006 20(8) 1488-1499. 

The dramatic effects observed when nitric oxide ( N=0) synthase is induced in cocultures of macrophages and tumor cells (Hibbs et al., 1987 Stuehr and Nathan, 1989) as well as Kupffer cells and hepatocytes (Billiar et al., 1989 ... 

According to the in vitro liver injury model, BCG-induced Kupffer cells are isolated from the livers of the rats injected via the tail vein with 3 mg of BCG 10 days before, and hepatocytes are isolated from the normal rats. The hepatocytes (1X 10 /mL), different concentrations of test compound, and BCG-induced Kupffer cells (1 X 10 /well) are cocultured for 48 h with 5-pg/mL LPS and the supernatants are collected. In a 96-well plate, 100pL of cell culture supernatant and 100pL of Griess reagent (10-g/L sulfanilamide and 1-g/L N-l-naphthylethylenediamine dihydrochloride in 2.5% phosphoric acid) are incubated at room temperature for lOmin. Absorbance at 540nm is measured. The nitrite concentration is calculated by comparing samples with standard solutions of sodium nitrite produced in the culture medium. 

Currently, it is clear that there is no in vitro model (and therefore no endpoint or marker) that can recapitulate in vitro, the role of the adaptive immune system in DILI (Uetrecht and Naisbitt, 2013). While there has been some progress at modeling the interactions of the innate immune system with hepatocytes through coculture with either monocytes or Kupffer cells, this is still a work in progress and needs to be refined based on further elucidation of immune mechanisms underlying human DILI. 

---