title: GSM7966243: Human, H. pylori-positive moderate intestinal metaplasia, scRNAseq, IM-HP-P_3_zhouyoulan; Homo sapiens; RNA-Seq
library_construction_protocol: The tissue samples were removed from patients and quickly immersed in Dulbecco's modified eagle medium (DMEM; Cat#11995065, Gibco) containing 10% fetal bovine serum (FBS; Cat#10091148, Gibco). Samples were minced into about 0.5 mm3 pieces and enzymatically digested in digestion medium DMEM supplemented with trypsin (Cat#15090046, Gibco), collagenase I (Cat# 17018029, Gibco), collagenase II (Cat# 17101015, Gibco) and collagenase IV (Cat# 17104019, Gibco). The samples were incubated and digested at 37℃ for approximately 60 min and then filtered using a 100μm strainer (Corning). Following centrifugation (300g) at 4℃ for 5 min, the cells were resuspended with medium containing red cell lysate (Cat#130094183, MACS). After standing 10min at 4℃, the single-cell suspension was centrifuged at 300g for 5min and resuspended in 100μl medium. Cell concentration and viability were detected under Luna Cell Counter. The suspensions were then converted to separate barcoded scRNA-seq libraries using the Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 (Cat#1000268, 10×genomics) according to the manufacturer's instruction. Libraries were sequenced on an Illumina Nova 6000 PE150