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The Isolation, Culture, and Propagation of Murine Intestinal Enteroids for the Study of Dietary Lipid Metabolism

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Organoids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1576))

Abstract

Since the initial report in 2009 by Sato and Clevers, primary enteroids have been of major interest in the fields of stem cell biology and gastrointestinal (GI) tract biology. More recently, we and others have made major inroads into the physiological relevance of these enteroid models and have shown that enteroids derived from the stomach, intestine, or colon recapitulate major functions of these tissues, namely, gastric acid secretion, lipid absorption and lipoprotein secretion, and ion transport. Here, we detail the isolation of stem cells from the small intestine and the culture and propagation of those stem cells into mature three-dimensional enteroids. We will also detail how we use enteroids to determine intestinal mechanisms behind dietary lipid absorption and lipoprotein secretion. The primary enteroid model is a powerful tool that significantly expands our ability to model GI tract function in vitro.

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Abbreviations

apo:

Apolipoprotein

BSA:

Bovine serum albumin

FFA:

Free fatty acids

ISCs:

Intestinal stem cells

LGR5:

Leucine-rich repeat-containing G-protein-coupled receptor 5

MG:

Monoacylglycerol

OA:

Oleic acid

TAG:

Triacylglycerol

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Correspondence to Alison B. Kohan .

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Li, D., Dong, H., Kohan, A.B. (2017). The Isolation, Culture, and Propagation of Murine Intestinal Enteroids for the Study of Dietary Lipid Metabolism. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2017_69

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  • DOI: https://doi.org/10.1007/7651_2017_69

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-7616-4

  • Online ISBN: 978-1-4939-7617-1

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