A Bacterial Infection Assay for Studying Anaerobic Bacteria-Host Cell Interactions
Published: November 30, 2023
Abstract
Source: Wunsch, C. M., et al. Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells. J. Vis. Exp. (2015)
This video demonstrates a technique for assessing the ability of Porphyromonas gingivalis, an anaerobic pathogenic bacterium, to adhere to and invade human umbilical vein endothelial cells (HUVECs). After infecting the host cells with the bacteria, the host cells are lysed to release both adhered and internalized bacteria. When the lysate is plated in an appropriate medium, the presence of colonies indicates successful bacteria-host cell interactions.
Protocol
The following protocols will describe methods for culturing and studying the invasion by the anaerobic species, P. gingivalis; however, these protocols may be used for a number of anaerobic pathogens. Although HUVECs are used, this protocol may be used for other eukaryotic cells both immune and non-immune. 1. Anaerobic Chamber Use and Maintenance Note: P. gingivalis is an anaerobe sensitive to normal levels …
Representative Results
Figure 1. Anaerobic vinyl chamber and its components. (A) A vinyl anaerobic chamber sealed completely from atmospheric oxygen provides workspace for two individuals at a time (32 in x 78 in). It contains an incubator set at 37 °C (back middle). (B) An airlock is used for the transfer of items from the lab environment to the anaerobic chamber. Pictured is an …
Disclosures
The authors have nothing to disclose.
Materials
| Vinyl Anaerobic Chamber-Type B | Coy Laboratory Products | Model 2000 incubator | |
| TSA II Trypticase Soy Agar w/5% Sheep Blood | BBL | 221261 | |
| Human Umbilical Vein Endothelial Cells 10-donor Pool | LifeLine Technology | FC-0044 | |
| VascuLife VEGF Medium Complete Kit | LifeLine Technology | LL-0003 | |
| TrypKit | LifeLine | LL-0013 | |
| Saponin | Riedel-de Haen | 16109 |
Cite This Article
A Bacterial Infection Assay for Studying Anaerobic Bacteria-Host Cell Interactions. J. Vis. Exp. (Pending Publication), e21795, doi: (2023).