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

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Citazione di questo articolo
Activation of Mouse Bone Marrow-Derived Macrophages in Response to Antibodies. J. Vis. Exp. (Pending Publication), e21606, doi: (2023).

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