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).