Functional assays with human patient-derived enteroid monolayers to assess the human gut barrier

Ibrahim M Sayed; Courtney Tindle; Ayden G Fonseca; Pradipta Ghosh; Soumita Das

doi:10.1016/j.xpro.2021.100680

Functional assays with human patient-derived enteroid monolayers to assess the human gut barrier

STAR Protoc. 2021 Jul 17;2(3):100680. doi: 10.1016/j.xpro.2021.100680. eCollection 2021 Sep 17.

Authors

Ibrahim M Sayed¹, Courtney Tindle^{2

3}, Ayden G Fonseca^{2

3}, Pradipta Ghosh^{2

3

4

5

6}, Soumita Das^{1

3

5

6}

Affiliations

¹ Department of Pathology, University of California, San Diego, CA 92093, USA.
² Department of Cellular and Molecular Medicine, University of California, San Diego, CA 92093, USA.
³ HUMANOID CoRE, University of California, San Diego, CA 92093, USA.
⁴ Department of Medicine, University of California, San Diego, CA 92093, USA.
⁵ Moore's Comprehensive Cancer Center, University of California, San Diego, CA 92093, USA.
⁶ Veterans Affairs Medical Center, VA San Diego Healthcare System, La Jolla, San Diego, CA 92093, USA.

Abstract

Here, we describe the use of polarized patient enteroid-derived monolayers (EDMs) to assess the impact of e-cigarettes on the human gut barrier. These EDMs can be adapted to culture in a 96-well plate for high-throughput screening. We model the effect of e-cigarettes by combining pathogens, enteroids, and e-cigarette vapor-infused media and assess gut barrier integrity, bacterial internalization, and inflammatory response of the gut epithelium. This protocol can be used to assess the effects of e-cigarette components on gut functions. For complete details on the use and execution of this protocol, please refer to Sharma et al. (2021).

Keywords: Cell Biology; Cell culture; High Throughput Screening; Immunology; Microbiology; Organoids; Stem Cells.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cell Culture Techniques / methods*
Epithelium
High-Throughput Screening Assays / methods
Humans
Intestinal Mucosa / drug effects*
Intestinal Mucosa / microbiology
Intestine, Small / cytology
Models, Biological
Organoids / cytology
Organoids / metabolism
Primary Cell Culture / methods
Vaping / adverse effects*

Abstract

Publication types

MeSH terms

Grants and funding