Cells and transfection
Primary mouse hepatocytes were isolated from BALB/c mice by following a
two-step perfusion protocol as previously described [16, 17], and
cultured in Hepato-Stim Hepatocyte Defined medium (BD Biosciences).
Primary human hepatocytes were cultured as previously described [18,
19]. IL-21 mRNA transfection was performed using TurboFect
Transfection Reagent (Thermo Fisher Scientific) according to the
manufacturer’s instructions. For LNP transfection, LNP particles were
diluted in serum free medium at a ratio of 1:5 and kept for 20 minutes
at room temperature, and then added into the cultured cells. Medium was
changed at 6-8 hours post transfection.