Assessing Internalized Porphyromonas gingivalis in Host Endothelial Cells
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)
In this video, we demonstrate an assay to assess the survival of Porphyromonas gingivalis inside human endothelial cells. The measure of survival and growth of internalized Porphyromonas gingivalis is evaluated by the number of colonies on the blood agar plate.
Protocol
1. Survival of internalized bacteria. Prepare P. gingivalis anaerobic bacteria until they reach mid-log growth (OD660 0.5-0.7). Centrifuge bacteria at 5,000 x g for 10 min. Place pelleted P. gingivalis back in the chamber, and discard the supernatant. Wash with PBS, and pellet bacteria again before resuspending in VEGF media. Prepare suspensions for all bacterial strains to be tested at OD660 of 0.7 which corresponds to the …
Divulgations
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 | |
| Gentamicin Sulfate Salt | Sigma-Aldrich | G-1264 | |
| Metronidazole | Sigma-Aldrich | M-3761 |
Citer Cet Article
Assessing Internalized Porphyromonas gingivalis in Host Endothelial Cells. J. Vis. Exp. (Pending Publication), e21799, doi: (2023).