Activation of Mouse Bone Marrow-Derived Macrophages in Response to Antibodies
Published: August 31, 2023
Abstract
Source: Kozicky, L. et al., Assessment of Antibody-based Drugs Effects on Murine Bone Marrow and Peritoneal Macrophage Activation. J. Vis. Exp. (2017)
In this video, we demonstrate the activation of bone marrow-derived macrophages in response to bacterial lipopolysaccharide and intravenous immunoglobulin.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Bone Marrow Macrophage Derivation and Activation with Antibodies Perform euthanasia using CO2 asphyxiation. Place the mouse in the induction chamber. Euthanize the mouse with 5% isoflurane anesthesia, for 60 – 90 s, until immobile and breathing is deep and slow. Turn off isoflurane anes…
Divulgazioni
The authors have nothing to disclose.
Materials
| Iscove's modified Dulbecco's medium (IMDM) | Life technologies | 12440053 | |
| Fetal Bovine Serum (FBS) | Life technologies | 12483-020 | |
| Recombinant murine macrophage colony-stimulating factor (MCSF) | Stemcell technologies | 78059 | |
| Penicillin-streptomycin | Life technologies | 15140148 | |
| Cell dissociation buffer | Life technologies | 13150016 | Enzyme-Free, Hanks's-based, EDTA |
| Lipopolysaccharide (LPS) | Sigma aldrich | L 4516 | From E. coli 0127:B8 |
| IVIg (Gammunex) | Grifols | Received from BC Children's Hospital, Transfusion Medicine | |
| IVIg (Gammagard liquid) | Baxter Healthcare Corporation | Received from BC Children's Hospital, Transfusion Medicine | |
| IVIg (Octagam) | Octapharma | Received from BC Children's Hospital, Transfusion Medicine | |
| Phosphate-buffered saline (PBS) (sterile), pH 7.4 | Life technologies | 10010023 | |
| Mouse IL-10 ELISA | BD biosciences | 555252 | |
| Mouse IL-12/23p40 ELISA | BD biosciences | 555165 | |
| 15 mL conical tube | BD biosciences | 352096 | |
| 50 mL conical tube | BD biosciences | 352070 | |
| Microcentrifuge tube (1.7 mL) | Diamed | SPE155-N | |
| 75 cm2 tissue culture treated flask | BD biosciences | 353136 | |
| Cell scraper | BD biosciences | 353085 |
Tags
Citazione di questo articolo
Activation of Mouse Bone Marrow-Derived Macrophages in Response to Antibodies. J. Vis. Exp. (Pending Publication), e21606, doi: (2023).