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