GSM9537956: FPC, 0.05% Chol, 32 weeks; Mus musculus; RNA-Seq
FNA procedure: FNA was performed with a 0.60 × 25 needle coupled to a 10 ml syringe, which was attached to a hand-grip syringe holder. The procedure consisted of the introduction of the needle into the tissue and suction while moving the needle back and forth repeatedly. Enough material was considered to have been obtained when the hub of the needle was filled with aspirate, then pressure applied on the syringe was released, and the needle is removed from the lesion. We harvested tissue from the 4 largest lobes of the liver (Right Lobe, Left Lobe, and 2 Middle Lobes). FNAs were placed into RPMI+10% FBS and kept on ice until processing. Cell dissociation: FNA biopsies were added to 10mL of liver digestion buffer pre-heated to 37°C. The liver digestion buffer was composed of 0.05% Collagenase type IV (Cat#17104-019, Gibco,) and 0.02% DNase (Cat#DN25, Sigma-Aldrich) in RPMI 1640 (Cat#22440, Invitrogen). Biopsies were dissociated into single-cell suspension for 15 minutes at 37°C on rocker. Red cells were removed using 1X BD Pharm Lyse™ Lysing Buffer (Cat#555899, BD Biosciences) and dead cells were removed using Dead Cell Removal Kit microbead solution (Cat#130–090-101, Miltenyi Biotech). Viable cells were particioned using 10x Chromium Controller and kit 3' v3.1. Library was performed according to the manufacter's instructions