GSM3140917: E17.5 1 SW; Mus musculus; RNA-Seq
Pancreata were isolated from mouse embryos and dissociated in TrypLE Express at 37C with pipet trituration at 5-minute intervals during incubation. E12.5 pancreata were dissociated for 10 minutes, E14.5 pancreata for 15 minutes, and E17.5 pancreata for 30 minutes. Dissociations were neutralized with FACS buffer (10% FBS + 2mM EDTA in phenol-red free HBSS). Dissociated cells were passed through a 30um cell strainer and stained with Sytox live/dead stain (Thermo Fisher), along with protocol specific stains noted in the individual sample extract protocol descriptions. Stained cells were washed twice in FACS buffer and then processed through a BD FACS Aria II. For Swiss Webster embryonic pancreatic dissociations, cells were also stained with the mesenchymal marker, CD140a. Stained cells were washed twice in FACS buffer and then processed through a BD FACS Aria II. To enrich for epithelial populations, live, CD140a- cells were collected from E12.5, E14.5, and E17.5 Swiss Webster pancreata. cDNA libraries were prepared for sequencing using standard protocol for the 10x Single Cell instrument with v2 kit and reagents. droplet-based single cell RNA-Seq
GEO Accession
GSM3140917