Journal Article

Controlling the polarity of human gastrointestinal organoids to investigate epithelial biology and infectious diseases

Monack, Denise M., Co, Julia Y., Amieva, Manuel R., Margalef-Català, Mar

2021

Overview

Human epithelial organoids—3D spheroids derived from adult tissue stem cells—enable investigation of epithelial physiology and disease and host interactions with microorganisms, viruses and bioactive molecules. One challenge in using organoids is the difficulty in accessing the apical, or luminal, surface of the epithelium, which is enclosed within the organoid interior. This protocol describes a method we previously developed to control human and mouse organoid polarity in suspension culture such that the apical surface faces outward to the medium (apical-out organoids). Our protocol establishes apical-out polarity rapidly (24–48 h), preserves epithelial integrity, maintains secretory and absorptive functions and allows regulation of differentiation. Here, we provide a detailed description of the organoid polarity reversal method, compatible characterization assays and an example of an application of the technology—specifically the impact of host–microbe interactions on epithelial function. Control of organoid polarity expands the possibilities of organoid use in gastrointestinal and respiratory health and disease research. The polarity of gastrointestinal organoids is reversed to study epithelial biology and host–microbe interactions. Access to the apical surface of the epithelium is increased while preserving epithelial integrity and secretory and absorptive functions.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/1647/1407/651 / 631/1647/767/2199 / 631/532/2437 / 631/80/85/2361 / Absorptivity / Analysis / Analytical Chemistry / Animals / Biological Techniques / Biology / Biomedical and Life Sciences / Cell culture / Cell Culture Techniques - methods / Cell Polarity / Communicable Diseases - microbiology / Computational Biology/Bioinformatics / Control / Coronaviruses / COVID-19 / Epithelial cells / Epithelial Cells - cytology / Epithelium / Experiments / Gastrointestinal system / Gastrointestinal Tract - cytology / Host-bacteria relationships / Humans / Infectious diseases / Integrity / Laboratories / Life Sciences / Mice / Microarrays / Microorganisms / Organic Chemistry / Organoids / Organoids - cytology / Pathogens / Physiological aspects / Polarity / Polarity (Biology) / Protocol / Severe acute respiratory syndrome coronavirus 2 / Spheroids / Stem cell transplantation / Stem cells / Surface chemistry / Suspension culture