GSM3178783: Patient 2 Total Liver Homogenate; Homo sapiens; RNA-Seq
The caudate lobe was cannulated with two or three 1.2-2mm diameter irrigation cannulae with olive tips, which were inserted into exposed vessels in the cut surface if the liver lobe (cannulae manufacturer: Ernst Kratz GmbH cat no: 1464LL, 1465LL). The lobe was flushed with HBS+EGTA at 4°C (perfusion rate of 10mL/min/cannulae), which removed any residual non-liver resident cells. Single cell isolation from the resected caudate liver lobe was performed with a modified  “two-step” collagenase procedure6,97 as described in Fig. 1b. Eight to 12 million viable cells were recovered per gram of tissue as determined by Beckman Vi-Cell trypan blue exclusion. Viability for TLHs by trypan blue exclusion varied between 50-90% viable for the five livers profiled. Caudate lobes weighed on average 30.2 g +/-SD of 8.367g. TLHs were taken directly after dissociation for scRNA Seq. 10X Genomics Single Cell 3' v2 Reagent Kit