GSM4983787: Day 85 OPC; Homo sapiens; RNA-Seq
Differentiating hOPC cultures were were dissociated into single cell suspension by incubating in accutase for ~45 minutes. The single cell suspension was then passed through a ~70 uM cell strainer. To purify PDGFRα expressing cells, the dissociated cells were resuspended in MACS buffer and CD90.2 (THY1.2) or O4 microbeads and incubated at room temperature for 15 minutes for cell binding. Please note that the hES cells used here are knock-in reporter cell lines where the PDGFRα drives the expression of tdTomato and CD90.2, allowing hES-derived OPCs to be purified using CD90.2 microbeads. After incubation with the beads, cells are generally run through the MS column twice without additional supplementation of MicroBeads to increase the purity and achieve ~90% tdTomato+ cells. The purified PDGFRα-tdTomato+ cells are then used to single cell capture by 10X. 10x Genomics Single Cell 3' v2 according to Manufacturers Recommendation