Single cell and bulk RNA sequencing of Enteroendocrine cells from human hormone reporter organoids. Single cell and bulk RNA sequencing of Enteroendocrine cells from human hormone reporter organoids

Enteroendocrine cell subpopulations were sorted by positivity for endogenous knock-in reporters in different hormones. Unbiased and biased FAC sorting was performed and coupled to single cell sorting by the SortSeq protocol (Muraro et al., 2016) and bulk sequencing. Overall design: Enteroendocrine cell differentiation was induced through overexpression of Neurogenin 3. After 5 days of differentiation, cells were collected by FACS and subjected to single cell and bulk RNA sequencing.肠内分泌细胞（Enteroendocrine cell）亚群通过针对不同激素的内源敲入报告基因（endogenous knock-in reporters）的阳性表达完成分选。研究开展了非偏向性与偏向性荧光激活细胞分选（FAC sorting），并通过SortSeq协议（SortSeq protocol，Muraro等人，2016）将该分选与单细胞分选流程相结合，同时配套批量测序。实验整体设计如下：通过过表达神经元素3（Neurogenin 3）诱导肠内分泌细胞分化；待分化培养满5天后，采用荧光激活细胞分选（FACS）收集细胞，随后开展单细胞及批量RNA测序。