A Simple and Efficient Monocyte Isolation Protocol from Human Peripheral Blood
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Copyright: © 2019 . This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Monocytes play an important role in immune regulation and are involved in a variety of effector functions. Isolation of human monocytes is essential when studying the functionality of these cells in health and disease. Once isolated, monocytes can then be cultured for functional assays or immunocytochemistry. However, the differentiation and activation state of human monocytes are affected by the culture conditions. Hence to overcome this, freshly isolated monocytes can be studied immediately after the isolation process via flow cytometry for immunophenotyping or functional assays acutely.
Our protocol isolates monocytes directly from human peripheral blood by negative selection. The kit used in our protocol removes non-monocytes with antibodies that recognize specific cell surface markers. Monocytes are not labelled with antibody and can easily be collected as a highly enriched population at the interface between the plasma and density gradient medium. Our protocol has a number of advantages: (1) isolated monocytes using our protocol are untouched and free of antibodies and (2) in comparison to other methodology for monocyte isolation, the technique presented in our protocol requires no specific equipment other than centrifuge.
