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Centrifugation

Centrifugation

Centrifugation is a separation technique based on differences in density or size. It is commonly used to separate solids from aqueous interferents. During centrifugation, the sample is placed in centrifugation tubes and spun at high angular velocity, which allows centrifugal force to act differentially on the different densities or masses of the components. After spinning, the supernatant liquid is decanted. Depending on the specific application, either the pellet or the supernatant is retained...

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Related Experiment Video

Updated: May 16, 2026

Isolation, Purification, and Differentiation of Osteoclast Precursors from Rat Bone Marrow

Isolation, Purification, and Differentiation of Osteoclast Precursors from Rat Bone Marrow

Published on: May 19, 2019

Density gradient centrifugation compromises bone marrow mononuclear cell yield.

Claudia Pösel¹, Karoline Möller, Wenke Fröhlich

¹Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany. claudia.poesel@izi.fraunhofer.de

|December 14, 2012

Summary

Ficoll-Paque density gradient centrifugation (DGC) significantly reduces bone marrow mononuclear cell (BMNC) yield and hematopoietic stem cell (HSC) content. Alternative methods like Percoll DGC or immunodepletion offer improved BMNC isolation for regenerative medicine.

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Last Updated: May 16, 2026

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Area of Science:

Cell Biology
Regenerative Medicine
Hematology

Background:

Bone marrow mononuclear cells (BMNCs) are crucial for regenerative medicine.
Current isolation methods, particularly Ficoll-Paque density gradient centrifugation (DGC), may compromise BMNC yield and function.
Understanding isolation impacts is vital for optimizing cell-based therapies.

Purpose of the Study:

To evaluate the effect of Ficoll-Paque DGC on BMNC yield and composition.
To compare Ficoll-Paque DGC with alternative isolation methods: adjusted Percoll DGC and immunomagnetic depletion of polymorphonuclear cells (PMNs).
To assess the impact on hematopoietic stem cell (HSC) content.

Main Methods:

BMNCs were isolated using Ficoll-Paque DGC, adjusted Percoll DGC, and immunomagnetic PMN depletion in an animal model.
BMNC recovery, subpopulation proportions, and colony-forming unit-granulocyte-macrophage (CFU-GM) for HSC content were analyzed.
A proof-of-concept study applied immunodepletion to human bone marrow aspirates.

Main Results:

Ficoll-Paque DGC resulted in maximal BMNC loss (lowest recovery: 25.6±5.8%).
Adjusted Percoll DGC and immunomagnetic PMN depletion showed significantly higher BMNC recovery (51.5±2.3% and 72.3±6.7%, respectively).
Hematopoietic stem cell (HSC) content was significantly reduced after Ficoll-Paque DGC compared to the other methods.

Conclusions:

Ficoll-Paque DGC significantly diminishes BMNC yield and HSC content.
Adjusted Percoll DGC and immunomagnetic PMN depletion are superior alternatives for isolating functional BMNCs.
Optimizing BMNC isolation protocols is essential for advancing preclinical and clinical regenerative medicine applications.