Chapter title |
Long-Term Culture of Intestinal Organoids
|
---|---|
Chapter number | 13 |
Book title |
Epithelial Cell Culture
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-8600-2_13 |
Pubmed ID | |
Book ISBNs |
978-1-4939-8599-9, 978-1-4939-8600-2
|
Authors |
Seung Bum Lee, Sung-Hoon Han, Sunhoo Park, Lee, Seung Bum, Han, Sung-Hoon, Park, Sunhoo |
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. |
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