A subscription to JoVE is required to view this content. Sign in or start your free trial.
An Assay to Study the Regulation of Macrophage Phagocytosis by Mesenchymal Stem Cells
Overview
This video demonstrates an assay to study mesenchymal stem cell (MSC) regulation of macrophage phagocytosis in a co-culture. Upon co-culturing MSC with macrophages, non-opsonized zymosan particles conjugated to a pH-sensitive fluorophore are added to be phagocytosed by the macrophages. With the increase in phagocytosis, the fluorescence intensity rises, allowing the determination of MSC-mediated regulation of phagocytic activity.
Protocol
NOTE: All medium preparation and cell culture techniques are carried out under aseptic conditions using a biosafety cabinet with laminar flow. All culture incubation steps described are carried out using an incubator designed to maintain an atmosphere of 37 °C, 5% CO2, and 95% humidity.
1. Cell culture
- Preparation of growth medium
- For MSC (mesenchymal stem cells) and LADMAC (mouse bone marrow-derived macrophages; it is d.......
Representative Results
Figure 1: Overview of the co-culture and phagocytosis assay workflow. An outline of the workflow for quantitative and qualitative analysis of MΦ phagocytosis activity in co-culture with MSC.
Supplementary File 1: A representative spreadsheet file of raw kinetic data from an optimal experiment.
This article has been published
Video Coming Soon
Source: Evans, J. F., et al. Mesenchymal Stem Cell Regulation of Macrophage Phagocytosis; Quantitation and Imaging. J. Vis. Exp. (2021).
ABOUT JoVE
Copyright © 2025 MyJoVE Corporation. All rights reserved