GSM7100143: Esophagus,GEJ and Stomach, from E15, E19, Pup and Adult, GEX; Mus musculus; RNA-Seq
Following gut extraction from prenatal (E15, E19) and postnatal (Pup, Adult) mice, the esophagus, gastroesophageal junction, and stomach were cut out. Tissue samples were washed in sterile PBS (Gibco, 14190-094) and minced with scissors. Minced tissue was processed separately by incubating in 0.5 mg ml−1 collagenase II (Calbiochem, 234155) in a shaker (45 min, 37 °C). Tissue and dissociated cells were pelleted (7 min, 1,000g, 4 °C), and the supernatant was discarded. The pellet was resuspended in  0.04% BSA-PBS solution and passed through a 40 µm cell strainer (BD Falcon, 352340). Cells from biological replicates were pooled and multiplexed based on the tissue region of origin and time point and library preparation for tissue were performed using 10x Genomics  3' CellPlex Kit and Single-Cell 3′ v3.1 RNA-seq kits according to manufacturer's protocol.