Establishment and characterization of adult human gastric epithelial progenitor‐like cell lines

作者：Yuan Gao, Ji Dong, Shuyue Qi, Xin Zhou, Xinglong Wu, Wendong Wang, Lu Wen, Wei Fu, Fuchou Tang · 发表于：Cell Proliferation · 年份：2022 · DOI：10.1111/cpr.13355 · 被引用次数：2 · 研究领域：Cancer Cells and Metastasis、Helicobacter pylori-related gastroenterology studies、Liver physiology and pathology

OBJECTIVE: Human gastric epithelial stem/progenitor cells are important for stomach homeostasis; however, the in vitro culture system of these cells remains immature. Although three-dimensional (3D) organoid culture has fundamentally changed the in vitro study of gastrointestinal tract, its use is limited by inaccessible luminal compartment, and difficulties of imaging and manipulation. To overcome these limitations of 3D organoid culture system, we established adult human gastric epithelial progenitor-like (hGEPL) cell lines using a novel robust monolayer cell culture system. MATERIALS AND METHODS: We established an in vitro gel-based monolayer culture system for normal human adult gastric epithelium, and compared it with traditional two-dimensional (2D) and 3D organoid culture systems using transcriptomics, immunofluorescence and cell viability experiments. At the same time, we used single-cell transcriptomics to compare the differences of the hGEPL cells in conditioned medium (Cond.) and in chemically defined medium (Chem.), the two most common media for organoid culture, in maintaining the stemness and proliferative activity of hGEPL cells. Finally, we explored the role of key niche factors in inducing hGEPL cell differentiation. RESULTS: The hGEPL cells were similar to the in vivo gastric epithelial stem/progenitor cells, which could stably proliferate in culture for a long time. Based on the established culture system, we explored signalling pathways that were important...