Isolation of Monocytes from Whole Blood by Immunomagnetic Negative Selection

Abstract

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

In this video, we demonstrate the isolation of monocytes from whole human blood by immunomagnetic negative separation. The isolated monocytes can be used for further downstream analysis.

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

1. Collect 40 mL of fresh, human whole blood (from either an HIV⁺ patient or healthy control) in four 10 mL ethylenediaminetetraacetic acid (EDTA) vacuum tubes (10 mL of blood per tube). Using sterile techniques under a biosafety cabinet, transfer all 40 mL of blood into one 50 mL conical propylene tube.
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Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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
PBS	Gibco	20012027
RPMI 1640 with L-Glutamine	Corning	10040CVMP