GSM5501835: Stomach epithelial organoid; Mus musculus; RNA-Seq
Esophageal and stomach organoids were harvested with ice-cold PBS, pelleted by centrifugation (5 Min, 300g, 4˚C), and Matrigel was removed. The process was repeated twice to remove Matrigel completely. Organoids were then incubated with warm TrypLE in a shaker (15 min, 37˚C, 180 rpm). Organoids were sheared using a 1 mL pipette by pipetting up and down 20 times. Dissociated cells were passed through a 40 µm cells strainer to obtain suspension of single cells, and the cells were washed with 0.1% BSA in 1XPBS.   Following the preparation of single-cell suspension, multiplexing of samples was performed according to the MULTI-seq protocol (McGinnis CS et al., Nat Methods. 2019). The barcoded samples were pooled together equally, and cell numbers were adjusted to 1000 cells/µl. A 10x Genomics Chromium™ Controller was used for partitioning single cells into nanoliter-scale Gel Bead-In-EMulsions (GEMs) and Single Cell 3′ v3.1 RNA-seq kit for reverse transcription, cDNA amplification and library construction (10xGenomics). The detailed protocol was provided by 10xGenomics. Chromium Next GEM Single Cell 3' v3.1 (10X Genomics)