Protocol for development, visualization, and quantification of mycobacterial biofilms on primary human airway epithelial cells

STAR Protocols, vol. 6, pp. 103872

Abstract

Tuberculosis (TB) displays several characteristics commonly linked to biofilm-associated infections, including recurrence of infection and resistance to antibiotics. Studying biofilm formation by mycobacteria on relevant mucosal surfaces advances our understanding of its role in TB pathogenesis and drug tolerance. Here, we present a protocol to promote biofilm formation by mycobacteria on differentiated primary pseudostratified human bronchial epithelial cell cultures. We then detail procedures to visualize and quantify biofilm matrix and biomass using electron and confocal microscopy and crystal violet staining. For complete details on the use and execution of this protocol, please refer to Barclay et al. 1

Authors 7

1. Amy M Barclay corresponding Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases (LUCID), Leiden University Medical Center (LUMC), 2333ZA Leiden, the Netherlands. Electronic address: a.m.de_waal@lumc.nl

2. Ronald W.A.L. Limpens Aachen Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Cell and Chemical Biology (CCB), LUMC, 2333ZA Leiden, the Netherlands

3. Montserrat Bárcena Aachen Cell and Chemical Biology

   Leiden University Medical Center

   Affiliation as printed

   Cell and Chemical Biology (CCB), LUMC, 2333ZA Leiden, the Netherlands

4. Tom H. M. Ottenhoff Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases (LUCID), Leiden University Medical Center (LUMC), 2333ZA Leiden, the Netherlands

5. Pieter Sicco Hiemstra Aachen PulmoScience Lab

   Leiden University Medical Center

   Affiliation as printed

   PulmoScience Lab, LUMC, 2333ZA Leiden, the Netherlands

6. Anne M. van der Does Aachen PulmoScience Lab

   Leiden University Medical Center

   Affiliation as printed

   PulmoScience Lab, LUMC, 2333ZA Leiden, the Netherlands

7. Simone A. Joosten corresponding Aachen

   Leiden University · Leiden University Medical Center

   Affiliation as printed

   Leiden University Center for Infectious Diseases (LUCID), Leiden University Medical Center (LUMC), 2333ZA Leiden, the Netherlands. Electronic address: s.a.joosten@lumc.nl