Isolation of Peripheral Blood Mononuclear Cells from Human Buffy Coats

Published: July 31, 2023

Abstract

Source: Mily, A., et al. Polarization of M1 and M2 Human Monocyte-Derived Cells and Analysis with Flow Cytometry upon Mycobacterium tuberculosis Infection. J. Vis. Exp. (2020)

In this video, we demonstrate the isolation of peripheral blood mononuclear cells, PBMCs, containing monocytes and lymphocytes from a human buffy coat by density gradient centrifugation. The isolated PBMCs can be used for further 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. Preparation of media NOTE: Details about all the reagents and consumables are provided in the Table of Materials. RPMI complete medium: Supplement RPMI 1640 with 1 mM sodium pyruvat…

Disclosures

The authors have nothing to disclose.

Materials

Lymphoprep Alere Technologies AS 11508545
RPMI 1640 Life Technologies Corporation SH30096.01
EDTA (0.5 M) Karolinska University hospital, Huddinge N/A
Na-pyruvate GE Healthcare Life Sciences SH300239.01
L-Glutamine GE Healthcare Life Sciences SH30034.01
HEPES GE Healthcare Life Sciences SH30237.01
Fetal bovine serum (FBS) Sigma-Aldrich F7524

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Cite This Article
Isolation of Peripheral Blood Mononuclear Cells from Human Buffy Coats. J. Vis. Exp. (Pending Publication), e21519, doi: (2023).

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