An Efficient Intestinal Organoid System of Direct Sorting to Evaluate Stem Cell Competition in Vitro.

ABSTRACT: Stem cell competition could shed light on the tissue-based quality control mechanism that prevents carcinogenesis. To quantitatively evaluate stem cell competition in vitro, we developed a two-color intestinal organoid forming system. First, we improved a protocol of culturing organoids from intestinal leucine-rich-repeat containing G-protein-coupled receptor 5 (Lgr5)- enhanced green fluorescent protein (EGFP)^high stem cells directly sorted on Matrigel without embedding. The organoid-forming potential (OFP) was 25% of Lgr5-EGFP^high cells sorted at one cell per well. Using this culture protocol with lineage tracing, we established a two-color organoid culture system by mixing stem cells expressing different fluorescent colors. To analyze stem cell competition, two-colo

SUBMITTER: Fujimichi Y 

PROVIDER: S-EPMC6937314 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature