Isolation of Mononuclear Cells (MNCs) from Blood Using Lymphoprep™ Density Gradient Media

Mononuclear cells (MNCs) are a diverse group of white blood cells encompassing lymphocytes (T cells, B cells, and NK cells), monocytes, and dendritic cells. These cell types play critical roles in the immune system, mediating both adaptive and innate immune responses.  Understanding MNC function requires their isolation from whole blood for further analysis. Lymphoprep (SKU:AXS-1114544-AXS-1115758) , a sterile, endotoxin-tested density gradient medium, provides a well-established and efficient method for achieving this goal.

image of Lymphoprep™

Principle and Advantages:

 Lymphoprep functions by exploiting differences in cell density. This sucrose-based solution possesses a specific density of 1.077 g/ml. When layered beneath a whole blood sample within a centrifuge tube, Lymphoprep creates a density gradient. During centrifugation, red blood cells (RBCs) and granulocytes (neutrophils, eosinophils, and basophils), having higher densities, sediment through the Lymphoprep layer and pellet at the bottom of the tube. Conversely, MNCs, with lower densities, migrate to the interface between the plasma layer (uppermost fraction) and the Lymphoprep layer. This distinct band of MNCs can be carefully aspirated for downstream applications.

Density Gradient Centrifugation

Several key advantages contribute to the widespread use of Lymphoprep™ for MNC isolation:

Rapidity: MNCs can be obtained within 15 minutes, facilitating efficient processing of samples.
Simplicity: The protocol involves minimal steps and requires basic laboratory equipment, making it accessible to researchers with varying levels of expertise.
Reproducibility: Lymphoprep™ has been a mainstay in cell isolation techniques for over four decades, consistently delivering reliable results.
Cost-effectiveness: Compared to other density gradient media, Lymphoprep offers a competitive cost option.
High Purity: The separation process yields highly purified MNC populations, minimizing contamination from other blood cell types.


MNCs isolated using Lymphoprep serve a multitude of purposes in research and clinical settings. Some prominent applications include:

Immunophenotyping: This technique allows for the identification and characterization of specific immune cell subsets within the MNC population using antibodies against cell surface markers.
Functional Assays: Isolated MNCs can be employed to assess their functional capabilities, such as proliferation in response to stimuli or cytokine production.
Cellular Therapies: MNCs, particularly T cells, are increasingly being explored for adoptive cell therapy, a promising approach for treating cancers and other immunological disorders.

In conclusion, Lymphoprep density gradient medium offers a rapid, simple, and reliable method for isolating highly purified MNCs from whole blood. This technique serves as a cornerstone for various immunological investigations and holds significant potential for advancing cellular therapy approaches.


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