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Methods Mol Biol. 2018;1817:123-135. doi: 10.1007/978-1-4939-8600-2_13.

Long-Term Culture of Intestinal Organoids.

Author information

1
Laboratory of Radiation Exposure & Therapeutics, National Radiation Emergency Medical Center, Korea Institute of Radiological and Medical Science, Seoul, Republic of Korea.
2
Laboratory of Radiation Exposure & Therapeutics, National Radiation Emergency Medical Center, Korea Institute of Radiological and Medical Science, Seoul, Republic of Korea. sunhoo@kirams.re.kr.

Abstract

The in vitro long-term expansion of primary intestinal epithelial cells has been hampered by the inability to maintain an immature stem cell population. Recent technical advances have led to the development of a novel in vitro culture system that can sustain intestinal stem cells (ISCs) using growth factors that mimic the intestinal microenvironment in combination with a three-dimensional (3D) culture. The resulting intestinal organoids display a crypt-villus architecture that recapitulates the native intestinal epithelium. Here, we describe our method for the long-term culture of intestinal epithelial organoids via consistent passaging using a gentle cell dissociation reagent to easily break the organoid into smaller pieces. The long-term cryopreservation and defining characteristics of these intestinal organoids also make this work relevant for the advancement of epithelial organoid-based therapeutic technologies by allowing the production of large numbers of cells for use in clinical applications.

KEYWORDS:

Crypt isolation; Intestinal epithelial cell; Intestine tissue; Lgr5; Long-term culture; Organoid

PMID:
29959709
DOI:
10.1007/978-1-4939-8600-2_13
[Indexed for MEDLINE]

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