GSM8489620: Myeloid cells, 5 weeks, CMO; Mus musculus; RNA-Seq
Perfused livers were disrupted using a gentleMACS tissue dissociator and resuspended in RPMI-1640 media containing 10% FBS. This suspension was centrifuged at 60 g for 1 min and the supernatant containing hepatic immune cells was collected and centrifuged again at 480 g for 8 min. The pellet was resuspended in 10 mL of 37.5% Percoll (Sigma) in PBS and centrifuged at 850 g for 30 min. The resulting pellet was washed followed by red blood cell lysis in ammonium-chloride-potassium (ACK) lysis buffer for 2 min at room temperature. Cells were loaded into the ports of a 10x-Genomics chip following a single-cell 3' kit. The gene expression and CMO libraries were sequenced using a Novaseq S4 chip (2x150bp PE).