Leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) has been identified as a marker of stem cells across multiple tissues. Lgr5-expressing cells are also regulators of tissue homeostasis and wound repair, and drivers of carcinogenic progression. The majority of information about Lgr5-expressing cells derives from genetically engineered mouse models. Human studies have been limited by a lack of specific reagents and experimental procedures for the purification of these cells. We recently demonstrated that antibody-based purification can be used to obtain viable LGR5-expressing cells from human primary tissues and patient derived organoids. Here, we provide detailed methods for the purification of these cells from colonic epithelial organoids generated from patient-derived tissues, from induced pluripotent stem cell (iPSC) derived intestinal organoids, and from freshly isolated patient tissue intestinal crypts. These methods will facilitate experimental analysis of human LGR5-expressing cells in development, wound healing, and cancer.
CITATION STYLE
Dame, M. K., Huang, S., Attili, D., Spence, J. R., & Colacino, J. A. (2020). Identification and isolation of human LGR5+ cells using an antibody-based strategy. In Methods in Molecular Biology (Vol. 2171, pp. 3–23). Humana Press Inc. https://doi.org/10.1007/978-1-0716-0747-3_1
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