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Smith NR, Swain JR, Davies PS, et al. Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations. Cell Mol Gastroenterol Hepatol. 2018;6(1):79-96doi: 10.1016/j.jcmgh.2018.02.007.
Smith, N. R., Swain, J. R., Davies, P. S., Gallagher, A. C., Parappilly, M. S., Beach, C. Z., Streeter, P. R., Williamson, I. A., Magness, S. T., & Wong, M. H. (2018). Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations. Cellular and molecular gastroenterology and hepatology, 6(1), 79-96. https://doi.org/10.1016/j.jcmgh.2018.02.007
Smith, Nicholas R, et al. "Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations." Cellular and molecular gastroenterology and hepatology vol. 6,1 (2018): 79-96. doi: https://doi.org/10.1016/j.jcmgh.2018.02.007
Smith NR, Swain JR, Davies PS, Gallagher AC, Parappilly MS, Beach CZ, Streeter PR, Williamson IA, Magness ST, Wong MH. Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations. Cell Mol Gastroenterol Hepatol. 2018 Mar 10;6(1):79-96. doi: 10.1016/j.jcmgh.2018.02.007. eCollection 2018. PMID: 29928673; PMCID: PMC6008251.
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Cell Mol Gastroenterol Hepatol. 2018 Mar 10;6(1):79-96. doi: 10.1016/j.jcmgh.2018.02.007. eCollection 2018.

Monoclonal Antibodies Reveal Dynamic Plasticity Between Lgr5- and Bmi1-Expressing Intestinal Cell Populations.

Cellular and molecular gastroenterology and hepatology

Nicholas R Smith, John R Swain, Paige S Davies, Alexandra C Gallagher, Michael S Parappilly, Catherine Z Beach, Philip R Streeter, Ian A Williamson, Scott T Magness, Melissa H Wong

Affiliations

  1. Department of Cell, Developmental and Cancer Biology, Oregon Health & Science University, Portland, Oregon.
  2. Department of Pediatrics, Oregon Health & Science University, Portland, Oregon.
  3. Oregon Health & Science University Stem Cell Center, Knight Cancer Institute, Oregon Health & Science University, Portland, Oregon.
  4. Department of Biomedical Engineering, Department of Medicine, Department of Cell Biology and Physiology, University of North Carolina, Chapel Hill, North Carolina.

PMID: 29928673 PMCID: PMC6008251 DOI: 10.1016/j.jcmgh.2018.02.007

Abstract

BACKGROUND & AIMS: Continual renewal of the intestinal epithelium is dependent on active- and slow-cycling stem cells that are confined to the crypt base. Tight regulation of these stem cell populations maintains homeostasis by balancing proliferation and differentiation to support critical intestinal functions. The hierarchical relation of discrete stem cell populations in homeostasis or during regenerative epithelial repair remains controversial. Although recent studies have supported a model for the active-cycling leucine-rich repeat-containing G-protein-coupled receptor 5 (Lgr5)

METHODS: We used novel monoclonal antibodies (mAbs) raised against murine intestinal epithelial cells in conjunction with ISC-green fluorescent protein (GFP) reporter mice to analyze relations between ISC populations by microscopy. Ex vivo 3-dimensional cultures, flow cytometry, and quantitative reverse-transcription polymerase chain reaction analyses were performed.

RESULTS: Two novel mAbs recognized distinct subpopulations of the intestinal epithelium and when used in combination permitted isolation of discrete Lgr5

CONCLUSIONS: These data showed the functional utility of murine mAbs in the isolation and investigation of Lgr5

Keywords: 3D, 3-dimensional; 4-OHT, 4-hydroxytamoxifen; APC, allophycocyanin; Bmi1; Bmi1, B lymphoma Mo-MLV insertion region 1 homolog; Egf, epidermal growth factor; FACS, fluorescence-activated cell sorting; GFP, green fluorescent protein; HBSS, Hank’s balanced salt solution; Hierarchy; ISC, intestinal stem cell; Intestinal Stem Cells; Lgr5; Lgr5, leucine-rich repeat-containing G-protein–coupled receptor 5; Lyz, lysozyme; OHSU, Oregon Health and Science University; PBS, phosphate-buffered saline; PE, Phycoerythrin; Plasticity; Rspo1, R-spondin1; TdT, tdTomato; Wnt, wingless-type MMTV (mouse mammary tumor virus) integration site; cDNA, complementary DNA; mAb, monoclonal antibody; mRNA, messenger RNA; qRT-PCR, quantitative reverse-transcription polymerase chain reaction

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