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Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy

The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem cells (ISCs) and progeny that progressively migrate and differentiate toward the tip of the villi. These processes, essential for gastrointestinal ho...

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Autores principales: Mahe, Maxime M., Sundaram, Nambirajan, Watson, Carey L., Shroyer, Noah F., Helmrath, Michael A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4401205/
https://www.ncbi.nlm.nih.gov/pubmed/25866936
http://dx.doi.org/10.3791/52483
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author Mahe, Maxime M.
Sundaram, Nambirajan
Watson, Carey L.
Shroyer, Noah F.
Helmrath, Michael A.
author_facet Mahe, Maxime M.
Sundaram, Nambirajan
Watson, Carey L.
Shroyer, Noah F.
Helmrath, Michael A.
author_sort Mahe, Maxime M.
collection PubMed
description The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem cells (ISCs) and progeny that progressively migrate and differentiate toward the tip of the villi. These processes, essential for gastrointestinal homeostasis, have been extensively studied using multiple approaches. Ex vivo technologies, especially primary cell cultures have proven to be promising for understanding intestinal epithelial functions. A long-term primary culture system for mouse intestinal crypts has been established to generate 3-dimensional epithelial organoids. These epithelial structures contain crypt- and villus-like domains reminiscent of normal gut epithelium. Commonly, termed “enteroids” when derived from small intestine and “colonoids” when derived from colon, they are different from organoids that also contain mesenchyme tissue. Additionally, these enteroids/colonoids continuously produce all cell types found normally within the intestinal epithelium. This in vitro organ-like culture system is rapidly becoming the new gold standard for investigation of intestinal stem cell biology and epithelial cell physiology. This technology has been recently transferred to the study of human gut. The establishment of human derived epithelial enteroids and colonoids from small intestine and colon has been possible through the utilization of specific culture media that allow their growth and maintenance over time. Here, we describe a method to establish a small intestinal and colon crypt-derived system from human whole tissue or biopsies. We emphasize the culture modalities that are essential for the successful growth and maintenance of human enteroids and colonoids.
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spelling pubmed-44012052015-04-24 Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy Mahe, Maxime M. Sundaram, Nambirajan Watson, Carey L. Shroyer, Noah F. Helmrath, Michael A. J Vis Exp Medicine The epithelium of the gastrointestinal tract is constantly renewed as it turns over. This process is triggered by the proliferation of intestinal stem cells (ISCs) and progeny that progressively migrate and differentiate toward the tip of the villi. These processes, essential for gastrointestinal homeostasis, have been extensively studied using multiple approaches. Ex vivo technologies, especially primary cell cultures have proven to be promising for understanding intestinal epithelial functions. A long-term primary culture system for mouse intestinal crypts has been established to generate 3-dimensional epithelial organoids. These epithelial structures contain crypt- and villus-like domains reminiscent of normal gut epithelium. Commonly, termed “enteroids” when derived from small intestine and “colonoids” when derived from colon, they are different from organoids that also contain mesenchyme tissue. Additionally, these enteroids/colonoids continuously produce all cell types found normally within the intestinal epithelium. This in vitro organ-like culture system is rapidly becoming the new gold standard for investigation of intestinal stem cell biology and epithelial cell physiology. This technology has been recently transferred to the study of human gut. The establishment of human derived epithelial enteroids and colonoids from small intestine and colon has been possible through the utilization of specific culture media that allow their growth and maintenance over time. Here, we describe a method to establish a small intestinal and colon crypt-derived system from human whole tissue or biopsies. We emphasize the culture modalities that are essential for the successful growth and maintenance of human enteroids and colonoids. MyJove Corporation 2015-03-06 /pmc/articles/PMC4401205/ /pubmed/25866936 http://dx.doi.org/10.3791/52483 Text en Copyright © 2015, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Medicine
Mahe, Maxime M.
Sundaram, Nambirajan
Watson, Carey L.
Shroyer, Noah F.
Helmrath, Michael A.
Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
title Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
title_full Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
title_fullStr Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
title_full_unstemmed Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
title_short Establishment of Human Epithelial Enteroids and Colonoids from Whole Tissue and Biopsy
title_sort establishment of human epithelial enteroids and colonoids from whole tissue and biopsy
topic Medicine
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4401205/
https://www.ncbi.nlm.nih.gov/pubmed/25866936
http://dx.doi.org/10.3791/52483
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