title: GSM5226815: Post-reperfusion liver sample [PR]; Homo sapiens; RNA-Seq
library_construction_protocol: The tissues were cut into 3 mm pieces, incubated with 1 mM EGTA Sigma-Aldrich, Cat. no. E0396-10G) and rotate at 37°C, 50 rpm for 10 minutes. After washing away the EGTA, the tissues were treated in the digestion solution (1 mg/ml Collagenase II + 1 mg/ml Collagenase IV + 50 ug/ml DNase I) at 100 rpm and 37 °C for 30 minutes. The cell suspension was passed through a 70 mm nylon cell strainer (BD, Cat. No. 352350, USA), and then centrifuged at 50×g for 3 minutes to collect the cell pellet, and then the remaining suspension centrifuged at 300×g for 5 minutes to collect the remaining cell pellet. Each tissue was resuspended to a concentration of 50-500 million cells per milliliter with resuspension buffer. Then used LIVE/DEAD Viability/Cytotoxicity Kit (Invitrogen, Cat. no. L3224, USA) to stained cells, and only AM+EH- cells were collected by FACS for each tissue Single-cell cDNA libraries were constructed using the standard procedure of GemCode technology (10X Genomics, USA)