Analysis of Metabolically Stressed Primary Hepatocytes with IL-1β Recombinant Protein for 24 Hours

RNA-seq analysis of primary hepatocytes isolated from C57BL/6N mice, stimulated with IL-1β recombinant protein to examine the transcriptional response to inflammatory signaling. Primary hepatocytes were isolated from C57BL/6N mouse livers. Liver perfusion was performed using 75 ml of an enzyme buffer solution containing collagenase type 1 (650 µg/ml; Worthington, Cat. LS004197) and collagenase P (50 µg/ml; Roche, Cat. 11213865001). Upon confirming successful cannulation, a cut was made at the hepatic portal vein for drainage of the perfusate. The liver was then dissected, finely minced, and filtered through a 100 µm filter. Hepatocytes were washed twice with enzyme buffer solution and subsequently resuspended in complete M199 medium. Cells were seeded in 12-well plates and incubated for 3 hours. Following this, the medium was changed, and cells were treated with IL-1β recombinant protein for 24 hours. RNA was isolated for sequencing from 2 untreated control samples (Control1, 2, 3) and 2 IL-1β treated samples (Treatment1, 2, 3).