In Vitro Culture and Differentiation of Primary Monocytes into Macrophages

Published: July 31, 2023

Abstract

Source: Plaisance-Bonstaff, K., et al. Isolation, Transfection, and Culture of Primary Human Monocytes. J. Vis. Exp. (2019)

This video demonstrates the priming of primary monocytes into macrophages. Primary human monocytes can be isolated and differentiated to an M1-like pro-inflammatory phenotype using the granulocyte-macrophage colony-stimulating factor (GM-CSF) or to an M2-like anti-inflammatory phenotype using the macrophage colony-stimulating factor (M-CSF).

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board. 1. Isolation of primary human monocytes by immunomagnetic negative selection Collect 40 mL of fresh, human whole blood (from either an HIV+ patient or healthy control) in four 10 mL ethylenediaminetetraacetic acid (EDTA) vacuum tu…

Disclosures

The authors have nothing to disclose.

Materials

0.5M EDTA Invitrogen AM9260G
BD Vacutainer Plastic Blood Collection Tubes with K2EDTA BD Biosciences 366643
Easy 50 EasySep Magnet StemCell Technologies 18002
Easy Sep Direct Human Monocyte Isolation Kit StemCell Technologies 19669
Fetal Bovin Serum Defined Heat Inactivated Hyclone SH30070.03HI
Nunc 35mm Cell Culture Dish Thermo Scientific 150318
PBS Gibco 20012027
Penicillin-Streptomycin Gibco 15140122
Recombinant Human GM-CSF R&D Systems 215-GM-050
Recombinant Human M-CSF R&D Systems 216-MC-025
RPMI 1640 with L-Glutamine Corning 10040CVMP

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Cite This Article
In Vitro Culture and Differentiation of Primary Monocytes into Macrophages. J. Vis. Exp. (Pending Publication), e21513, doi: (2023).

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