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Related Concept Videos

Overview Of Cell Separation And Isolation01:20

Overview Of Cell Separation And Isolation

Cell separation was first achieved in 1964 by S. H. Seal, who separated large tumor cells from the smaller blood cells using filtration. Two years later, Pohl and Hawk performed experiments on how cells respond differently to a nonuniform electric field based on the cell type. Such observations were the inception of cell separation methods, which allow isolating a single cell type from a heterogeneous sample.
Subcellular Fractionation01:32

Subcellular Fractionation

The homogenate obtained after cell lysis contains various membrane-bound organelles that can be further separated into pure fractions by subcellular fractionation. These isolates are used to study specific cellular components, analyze localized protein activity, and are even employed in diagnostics. Fractionation is typically achieved using centrifugation methods, the most common being density-gradient and differential centrifugation.
Differential Centrifugation
Differential centrifugation is...

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Related Experiment Video

Updated: May 19, 2026

Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation
07:11

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Published on: January 25, 2016

Discontinuous density gradient separation of human mononuclear leucocytes using Percoll as gradient medium.

A J Ulmer, H D Flad

    Journal of Immunological Methods
    |January 1, 1979
    PubMed
    Summary

    Percoll medium effectively separates human mononuclear leukocytes by density, yielding pure monocyte and lymphocyte populations. This method enhances cell isolation for immunological studies, with specific fractions showing optimal responses in various assays.

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    Area of Science:

    • Immunology
    • Cell Biology
    • Hematology

    Background:

    • Accurate isolation of human mononuclear leukocytes is crucial for immunological research.
    • Existing cell fractionation methods can be labor-intensive and yield variable purity.

    Purpose of the Study:

    • To evaluate the efficacy of Percoll as a commercially available medium for human mononuclear leukocyte fractionation.
    • To assess the reproducibility and purity of cell populations obtained using Percoll gradients.

    Main Methods:

    • Human mononuclear leukocytes were fractionated using density gradients of Percoll.
    • Separated cell populations were analyzed for purity using morphological and functional criteria.
    • Cellular responses were assessed via spontaneous DNA synthesis, mixed leukocyte culture (MLC), and mitogen stimulation (PHA, PWM).

    Main Results:

    • Percoll gradients achieved high reproducibility in cell fractionation based on density.
    • Monocytes were isolated with 70%-90% purity in low-density fractions.
    • Lymphocytes were isolated with up to 99% purity in high-density fractions.
    • Specific density interfaces showed optimal responses for spontaneous DNA synthesis, MLC, PHA, and PWM stimulation.
    • Colony-forming myelopoietic stem cells and T-lymphocytes were found in low-density fractions.

    Conclusions:

    • Percoll is a reliable and efficient medium for separating human mononuclear leukocytes.
    • The method allows for the isolation of distinct monocyte and lymphocyte populations with high purity.
    • Fractionation by Percoll density can yield cells with specific functional properties, valuable for immunological assays.