Pluripotent Stem Cell-Derived Gastric Lineage

The global population infected with Helicobacter pylori is enormous, and gastrointestinal diseases including gastritis, peptic ulcers, gastric antral dysplasia, and gastric cancer occur at high incidence, with lesions frequently concentrated in the gastric antrum. Current gastric disease research faces significant technical challenges: conventional animal models such as mice and rats exhibit substantial differences from humans in gastric embryonic development, epithelial cell subtype composition, and pathological responses to H. pylori, making it difficult to accurately reproduce human gastric physiology and disease progression. In addition, clinical acquisition of primary human gastric mucosal tissue is challenging, and these cells rapidly lose functional characteristics in vitro, preventing long-term stable culture. These limitations greatly restrict investigations into gastric disease mechanisms, antibacterial/gastroprotective/anticancer drug development, and gastric mucosal regeneration technologies.

In 2014, a landmark study published in Nature established the first standardized stepwise differentiation system for the stable generation of human gastric organoids (hGOs), faithfully recapitulating the entire process of human gastric embryonic development. Using human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) as starting materials, this protocol precisely regulates multiple key developmental signaling pathways, including Activin A, Wnt, FGF, BMP, retinoic acid (RA), and EGF, through four major differentiation stages. First, highly purified definitive endoderm is induced; subsequently, Noggin is used to inhibit hindgut fate, while retinoic acid activates the gastric-specific transcription factor PDX1 to generate posterior foregut gastric progenitors. These progenitors are then transferred into long-term three-dimensional extracellular matrix culture, where stage-specific EGF concentration regulation promotes organoid expansion and terminal maturation of endocrine lineages. The entire differentiation process requires approximately 34 days. Mature gastric antrum organoids develop folded, gland-like three-dimensional structures and contain major gastric cell populations, including surface mucous cells, neck mucous cells, SOX9-positive proliferative progenitor cells, LGR5-positive gastric stem cells, and all major gastric antral endocrine subtypes, including gastrin-, somatostatin-, ghrelin-, and serotonin-producing cells. These organoids lack gastric corpus parietal cell contamination, intestinal, or pancreatic ectopic lineages, and their transcriptional profiles closely match those of human fetal gastric antrum tissue. Studies have further demonstrated the dual regulatory role of EGF signaling: high concentrations of EGF promote organoid growth and glandular morphogenesis, whereas low concentrations of EGF relieve NEUROG3 suppression to drive endocrine cell differentiation. Meanwhile, standardized Helicobacter pylori infection models established using these organoids successfully reproduce human-specific early pathological responses, including CagA toxin binding to the c-Met receptor and induction of abnormal epithelial proliferation, accurately modeling gastric infection-associated pathogenic pathways. This differentiation platform features precisely controlled signaling timing, high reproducibility, and scalability. Based on this human gastric antrum organoid system, applications include H. pylori-associated gastric disease modeling, high-throughput screening of gastrointestinal mucosal protective, acid-suppressive, and anticancer drugs, studies of human gastric embryonic development and gastric stem cell biology, and development of regenerative therapies for gastric mucosal injury. This platform provides a high-fidelity, traceable, standardized human gastric tissue model for digestive disease research and preclinical drug evaluation.

The differentiation process of pluripotent stem cell-derived gastric lineage. Through a four-stage serum-free directed induction system, pluripotent stem cells sequentially undergo definitive endoderm, foregut gastric progenitor, gastric organoid expansion, and endocrine maturation stages, forming human gastric antrum organoids with gastric gland-like structures, multiple gastric epithelial cell types, and endocrine cells within 34 days. This platform can simulate Helicobacter pylori infection and gastric development processes, providing a high-fidelity humanized model for gastric disease mechanism studies, drug screening, gastric mucosal repair, and regenerative medicine research.