A Fully Differentiated Macrophage Infection Model of Monocyte-Derived Cells with Mycobacterium tuberculosis
Published: September 29, 2023
Abstract
Source: Mily, A., et al. Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection. J. Vis. Exp.(2020).
This video demonstrates a method to study the responses to Mycobacterium tuberculosis infection in human M1 and M2 polarized monocyte-derived macrophages. The use of flow cytometry enables the study of a variety of surface markers associated with infected and uninfected macrophage populations.
Protocol
1. Mtb infection of monocyte-derived cells NOTE: The following steps must be performed in a BSL-3 facility. Resuspend the Mycobacterium tuberculosis (Mtb) pellet in serum-free RPMI medium in a new sterile 50 mL tube and adjust the final bacterial concentration to approximately 5 x 106 CFU/mL. Remove the cell culture medium from the 6-well plate(s) containing monocyte-derived cells. Add 1 mL of serum-free RPMI medium to…
Divulgations
The authors have nothing to disclose.
Materials
| 8-well chamber slides | Lab-Tek | 154534 | |
| RPMI 1640 | Life Technologies Corporation | SH30096.01 | |
| IFN-γ | Peprotech | 300-02 | |
| Na-pyruvate | GE Healthcare Life Sciences | SH300239.01 | |
| L-Glutamine | GE Healthcare Life Sciences | SH30034.01 | |
| Falcon 6-well Flat Bottom plates | Corning Life Sciences | 353046 | |
| Fetal bovine serum (FBS) | Sigma-Aldrich | F7524 | |
| GM-CSF | Peprotech | 300-03 |
Tags
Citer Cet Article
A Fully Differentiated Macrophage Infection Model of Monocyte-Derived Cells with Mycobacterium tuberculosis. J. Vis. Exp. (Pending Publication), e21694, doi: (2023).