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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
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
References
Mansbach CM 2nd, Gorelick F (2007) Development and physiological regulation of intestinal lipid absorption. II. Dietary lipid absorption, complex lipid synthesis, and the intracellular packaging and secretion of chylomicrons. Am J Physiol Gastrointest Liver Physiol 293:G645–G650
Date S, Sato T (2015) Mini-gut organoids: reconstitution of the stem cell niche. Annu Rev Cell Dev Biol 31:269–289
Wilson TH, Wiseman G (1954) The use of sacs of everted small intestine for the study of the transference of substances from the mucosal to the serosal surface. J Physiol 123:116–125
Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE, van Es JH, Abo A, Kujala P, Peters PJ et al (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459:262–265
VanDussen KL, Carulli AJ, Keeley TM, Patel SR, Puthoff BJ, Magness ST, Tran IT, Maillard I, Siebel C, Kolterud Å et al (2012) Notch signaling modulates proliferation and differentiation of intestinal crypt base columnar stem cells. Development 139:488–497
Murphy CL, Polak JM (2002) Differentiating embryonic stem cells: GAPDH, but neither HPRT nor beta-tubulin is suitable as an internal standard for measuring RNA levels. Tissue Eng 8:551–559
Fevr T, Robine S, Louvard D, Huelsken J (2007) Wnt/beta-catenin is essential for intestinal homeostasis and maintenance of intestinal stem cells. Mol Cell Biol 27:7551–7559
Tetteh PW, Basak O, Farin HF, Wiebrands K, Kretzschmar K, Begthel H, Van Den Born M, Korving J, De Sauvage F, Van Es JH et al (2016) Replacement of lost Lgr5-positive stem cells through plasticity of their enterocyte-lineage daughters. Cell Stem Cell 18:203–213
Levy E, Beaulieu JF, Delvin E, Seidman E, Yotov W, Basque JR, Ménard D (2000) Human crypt intestinal epithelial cells are capable of lipid production, apolipoprotein synthesis, and lipoprotein assembly. J Lipid Res 41:12–22
Foulke-Abel J, In J, Yin J, Zachos NC, Kovbasnjuk O, Estes MK, de Jonge H, Donowitz M (2016) Human enteroids as a model of upper small intestinal ion transport physiology and pathophysiology. Gastroenterology 150:638–649.e8
Mahe MM, Sundaram N, Watson CL, Shroyer NF, Helmrath MA (2015) Establishment of human epithelial enteroids and colonoids from whole tissue and biopsy. J Vis Exp 97:e52483–e52496
Chateau D, Pauquai T, Delers F, Rousset M, Chambaz J, Demignot S (2005) Lipid micelles stimulate the secretion of triglyceride-enriched apolipoprotein B48-containing lipoproteins by Caco-2 cells. J Cell Physiol 202:767–776
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media New York
About this protocol
Cite this protocol
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
Download citation
DOI: https://doi.org/10.1007/7651_2017_69
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-7616-4
Online ISBN: 978-1-4939-7617-1
eBook Packages: Springer Protocols