Quantification of Bacterial Infection in Host Cells by the Colony-Forming Unit Assay
Published: September 29, 2023
Abstract
Source: Gayle, P. et al., Using a Bacterial Pathogen to Probe for Cellular and Organismic-level Host Responses. J. Vis. Exp. (2019).
This video demonstrates an in vitro quantification technique to determine Listeria monocytogenes infection of macrophages by the colony-forming unit assay. The bacterial colonies on the agar plate represent the intracellular bacterial population that escapes the host-defense mechanism and survives within the macrophages.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Preparing Cells for Infection Propagate mouse macrophage-like cell line RAW 264.7 in DMEM tissue culture media supplemented with 10% fetal calf serum (henceforth referred to as DMEM/FCS) using a 125 mL flask and a tissue culture incubator maintained at 37 °C/5% CO2. The day before the infection…
Disclosures
The authors have nothing to disclose.
Materials
| BHI (Brain Heart Infusion) broth | EMD Milipore | 110493 | |
| DMEM media | Gibco | 11965-092 | |
| FBS – Heat-Inactivated | Sigma-Aldrich | F4135-500ML | |
| LB agar | Grow Cells MS | MBPE-4040 | |
| TPP Tissue Culture 48-Well Plates | MIDSCI | TP92048 |
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Cite This Article
Quantification of Bacterial Infection in Host Cells by the Colony-Forming Unit Assay. J. Vis. Exp. (Pending Publication), e21647, doi: (2023).