Format

Send to

Choose Destination
Methods Mol Biol. 2016;1422:33-40. doi: 10.1007/978-1-4939-3603-8_4.

An Air-Liquid Interface Culture System for 3D Organoid Culture of Diverse Primary Gastrointestinal Tissues.

Author information

1
Hematology Division, Department of Medicine, Stanford University School of Medicine, 265 Campus Drive, Stanford, CA, 94305, USA.
2
Hematology Division, Department of Medicine, Stanford University School of Medicine, 265 Campus Drive, Stanford, CA, 94305, USA. cjkuo@stanford.edu.

Abstract

Conventional in vitro analysis of gastrointestinal epithelium usually relies on two-dimensional (2D) culture of epithelial cell lines as monolayer on impermeable surfaces. However, the lack of context of differentiation and tissue architecture in 2D culture can hinder the faithful recapitulation of the phenotypic and morphological characteristics of native epithelium. Here, we describe a robust long-term three-dimensional (3D) culture methodology for gastrointestinal culture, which incorporates both epithelial and mesenchymal/stromal components into a collagen-based air-liquid interface 3D culture system. This system allows vigorously expansion of primary gastrointestinal epithelium for over 60 days as organoids with both proliferation and multilineage differentiation, indicating successful long-term intestinal culture within a microenvironment accurately recapitulating the stem cell niche.

KEYWORDS:

Air–liquid interface; Gastrointestinal tissue culture; Organoid; Stem cell niche; Three dimensional culture

PMID:
27246020
DOI:
10.1007/978-1-4939-3603-8_4
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center