Title: Identification and Isolation of Human LGR5+ Cells Using an Antibody-Based Strategy.

Authors: Dame, Michael K; Huang, Sha; Attili, Durga; Spence, Jason R; Colacino, Justin A

Published In Methods Mol Biol, (2020)

Abstract: 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.

PubMed ID: 32705632

MeSH Terms: Animals; Cells, Cultured; Colon/cytology*; Flow Cytometry; Humans; Induced Pluripotent Stem Cells/cytology; Induced Pluripotent Stem Cells/metabolism; Mice; Organoids/cytology*; Organoids/metabolism; Receptors, G-Protein-Coupled/genetics; Receptors, G-Protein-Coupled/metabolism*; Stem Cells/cytology*; Stem Cells/metabolism