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Development. 2018 Mar 14;145(6). pii: dev153049. doi: 10.1242/dev.153049.

Identification, isolation and characterization of human LGR5-positive colon adenoma cells.

Author information

1
Department of Internal Medicine, Division of Gastroenterology, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
2
Department of Pathology, University of Michigan, Ann Arbor, MI 48109, USA.
3
Department of Surgery, University of Michigan, Ann Arbor, MI 48109, USA.
4
Miltenyi Biotec GmbH, Bergisch Gladbach, 51429, Germany.
5
Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA.
6
Department of Molecular and Integrative Physiology, University of Michigan, Ann Arbor, MI 48109, USA.
7
Department of Research and Development, Progenity, Inc., Ann Arbor, MI 48109, USA.
8
Cellular and Molecular Biology Program, University of Michigan, Ann Arbor, MI 48109, USA.
9
BioCentury Publications, Redwood City, CA 94065, USA.
10
Department of Internal Medicine, Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, MI 48109, USA.
11
Department of Pharmacology, University of Michigan, Ann Arbor, MI 48109, USA.
12
Department of Environmental Health Sciences, University of Michigan School of Public Health, Ann Arbor, MI 48109, USA colacino@umich.edu.
13
Department of Nutritional Sciences, University of Michigan School of Public Health, Ann Arbor, MI 48109, USA.

Abstract

The intestine is maintained by stem cells located at the base of crypts and distinguished by the expression of LGR5. Genetically engineered mouse models have provided a wealth of information about intestinal stem cells, whereas less is known about human intestinal stem cells owing to difficulty detecting and isolating these cells. We established an organoid repository from patient-derived adenomas, adenocarcinomas and normal colon, which we analyzed for variants in 71 colorectal cancer (CRC)-associated genes. Normal and neoplastic colon tissue organoids were analyzed by immunohistochemistry and fluorescent-activated cell sorting for LGR5. LGR5-positive cells were isolated from four adenoma organoid lines and were subjected to RNA sequencing. We found that LGR5 expression in the epithelium and stroma was associated with tumor stage, and by integrating functional experiments with LGR5-sorted cell RNA sequencing data from adenoma and normal organoids, we found correlations between LGR5 and CRC-specific genes, including dickkopf WNT signaling pathway inhibitor 4 (DKK4) and SPARC-related modular calcium binding 2 (SMOC2). Collectively, this work provides resources, methods and new markers to isolate and study stem cells in human tissue homeostasis and carcinogenesis.

KEYWORDS:

OLFM4; Organoid; SMOC2; Stem cell; Stroma; Wnt signaling

PMID:
29467240
PMCID:
PMC5897593
[Available on 2019-03-15]
DOI:
10.1242/dev.153049
[Indexed for MEDLINE]
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Conflict of interest statement

Competing interestsO.H., D. Agorku and A.B. are full-time employees of Miltenyi Biotec. J.L. and J.A.S. are full-time employees of Progenity, Inc. The remaining co-authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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