Glucagon-like peptide-1 (GLP-1) from intestinal L-cells stimulates insulin secretion and reduces appetite after food ingestion, and it is the basis for drugs against type-2 diabetes and obesity. Drugs targeting L- and other enteroendocrine cells are under development, with the aim to mimic endocrine effects of gastric bypass surgery, but they are difficult to develop without human L-cell models. Human ileal organoids, engineered by CRISPR-Cas9, express the fluorescent protein Venus in the proglucagon locus, enabling maintenance of live, identifiable human L-cells in culture. Fluorescence-activated cell sorting (FACS)-purified organoid-derived L-cells, analyzed by RNA sequencing (RNA-seq), express hormones, receptors, and ion channels, largely typical of their murine counterparts. L-cells are electrically active and exhibit membrane depolarization and calcium elevations in response to G-protein-coupled receptor ligands. Organoids secrete hormones in response to glucose and other stimuli. The ability to label and maintain human L-cells in organoid culture opens avenues to explore L-cell function and develop drugs targeting the human enteroendocrine system.

摄入食物后，来自肠道L细胞的胰高血糖素样肽1（GLP-1）刺激胰岛素分泌并降低食欲，它是抗2型糖尿病和肥胖症药物的基础。针对L细胞和其他肠内分泌细胞的药物正在开发中，目的是模仿胃搭桥手术的内分泌作用，但是如果没有人类L细胞模型，则很难开发它们。由CRISPR-Cas9设计的人类回肠类器官在前胰高血糖素基因座中表达荧光蛋白维纳斯，从而能够在培养物中维持活的，可识别的人类L细胞。荧光激活细胞分选（FACS）纯化的类器官衍生L细胞，通过RNA测序（RNA-seq）分析，表达激素，受体和离子通道，这是它们鼠类对应物的典型特征。L细胞具有电活性，并响应于G蛋白偶联的受体配体表现出膜去极化和钙升高。响应于葡萄糖和其他刺激，类器官分泌激素。在类器官培养物中标记和维持人类L细胞的能力为探索L细胞功能和开发针对人类肠内分泌系统的药物开辟了道路。