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

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Description

<jats:title>Abstract</jats:title><jats:p>Stem cell competition could shed light on the tissue-based quality control mechanism that prevents carcinogenesis. To quantitatively evaluate stem cell competition <jats:italic>in vitro</jats:italic>, 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)<jats:sup>high</jats:sup> stem cells directly sorted on Matrigel without embedding. The organoid-forming potential (OFP) was 25% of Lgr5-EGFP<jats:sup>high</jats:sup> 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-color organoids were formed by mixing X-ray-irradiated and non-irradiated intestinal stem cells. In the two-color organoids, irradiated stem cells exhibited a growth disadvantage, although the OFP of irradiated cells alone did not decrease significantly from that of non-irradiated cells. These results suggest that stem cell competition can be evaluated quantitively <jats:italic>in vitro</jats:italic> using our new system.</jats:p>

Journal

  • Scientific Reports

    Scientific Reports 9 (1), 20297-, 2019-12-30

    Springer Science and Business Media LLC

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