title: GSM3535278: AXLN3; Homo sapiens; RNA-Seq
library_construction_protocol: Immediately after surgery, human LNs were transferred into RPMI on ice and cut into small pieces, and digested for 1h with RPMI containing collagenase P, dispase and DNase. Hematopoietic cells were depleted with EasySepTM Human CD45 Depletion Kit (Stem Cell Technologies), and the single cell suspension was incubated with PE-anti-PDPN, AF488-anti-CD45, APC-anti-CD31 and LIVE/DEADTM Fixable Near-IR Dead Cell Stain kit (ThermoFisher Scientific, L10119) for 30 min. Live CD45- PDPN+ CD31+ cells known to include LECs were sorted with FACS Aria (BD Bioscience) or SH800S (Sony) into RPMI medium containing 10% FCS. Sorted cells were immediately count and run on the 10X Chromium (10X Genomics). ScRNAseq libraries were prepared according to the manufacturer's instructions (CG00052 RevB) using Chromium™ Single Cell 3' Library and Gel Bead Kit v2 (10X Genomics, 120237) and Chromium™ Single Cell A Chip Kit (10X Genomics, 120236).
identifiers.namespace: GEO
identifiers.id: GSM3535278