Single cell RNAseq of small intestinal stromal cells

We used scRNAseq approaches (Smartseq2 and 10X Genomics) to investigate the diversity of small intestinal stromal cells. Small intestines from adult C57Bl/6 mice were harvested and digested with Liberase TL. Single, live, CD45neg, EpCAMneg, CD31neg, gp38+ cells were either sorted into 384-well plates (Smartseq2) or captured in an oil emulsion (10X Genomics). RNA was amplified, barcoded and sequenced.本研究采用单细胞RNA测序（single-cell RNA sequencing, scRNAseq）技术，依托Smart-seq2与10X Genomics两种实验平台，探究小肠基质细胞的异质性。研究首先取材成年C57Bl/6小鼠的小肠组织，经Liberase TL酶消化后，分选出单细胞、活体细胞、CD45阴性、EpCAM阴性、CD31阴性且gp38阳性的细胞群；将上述细胞群分别接种至384孔板以开展Smart-seq2实验，或通过油滴乳剂体系完成细胞捕获以适配10X Genomics平台。随后对RNA进行扩增、添加条形码标记并完成测序。