GSM4411627: RP009; Homo sapiens; RNA-Seq
Monocyte preparation uses a modification of published protocols. Briefly, ~20 ml blood drawn in sodium heparin was processed immediately in the lab in the Clinical Research Center at Columbia University. PBMCs were isolated by gradient Ficoll paque centrifugation, which maintains cell viability and prevents ex vivo activation during cell recovery. Cells were stained with antibodies against human HLADR, CD14 and CD16 and monocyte subsets defined as HLADR+CD14++CD16-(classical), HLADR+CD14++CD16+ (intermediate), HLADR+CD14dim/CD16++ (nonclassical, patrolling monocyte). DAPI staining was used to exclude dead cells. Monocytes were sorted by a BD Influx Sorter into tubes for real-time 10x Genomics analysis. Samples were prepared using the 10x Genomics Chromium Single Cell 3' Reagent Kits v2 according to manufacturer's instructions.12 cycles for cDNA amplification and 12 cycles for sample index PCR were used.
GEO Accession
GSM4411627