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Related Experiment Video

Updated: Apr 1, 2026

Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix
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Isolation of Human Mesenchymal Stem Cells and their Cultivation on the Porous Bone Matrix

Published on: February 9, 2015

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Easily-handled method to isolate mesenchymal stem cells from coagulated human bone marrow samples.

Heng-Xiang Wang1, Zhi-Yong Li1, Zhi-Kun Guo1

  • 1Heng-Xiang Wang, Department of Hematology, General Hospital of Air Force, Beijing 100042, China.

World Journal of Stem Cells
|October 6, 2015
PubMed
Summary

Urokinase treatment effectively isolates mesenchymal stem cells (MSCs) from clotted bone marrow, yielding cell numbers comparable to fresh samples. This method offers a viable solution for processing difficult-to-handle, coagulated human bone marrow specimens.

Keywords:
Coagulated marrow sampleIsolationMesenchymal stem cellsUrokinase

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Regenerative Medicine

Background:

  • Mesenchymal stem cells (MSCs) are crucial for regenerative medicine.
  • Bone marrow aspiration can result in sample coagulation, hindering MSC isolation.
  • Efficient MSC isolation from coagulated samples is challenging.

Purpose of the Study:

  • To develop a simple and effective method for isolating MSCs from coagulated human bone marrow.
  • To compare the efficacy of different pre-treatment methods on clot samples.

Main Methods:

  • Human bone marrow samples were treated with thrombin to induce coagulation.
  • Coagulated samples were pre-treated with urokinase, mechanical cutting, or left untreated.
  • Un-coagulated samples served as controls.
  • Colony-forming unit-fibroblast (CFU-F) numbers were quantified.
  • Isolated cells were characterized by flow cytometry and in vitro differentiation assays.

Main Results:

  • Urokinase treatment yielded significantly higher CFU-F numbers compared to mechanical cutting or untreated clots.
  • CFU-F counts in urokinase-treated samples were comparable to un-coagulated controls.
  • Isolated cells exhibited typical MSC surface markers (CD44, CD73, CD90) and differentiation potential.
  • Storage at 4°C reduced CFU-F counts, but urokinase treatment maintained higher yields.

Conclusions:

  • Urokinase pre-treatment is an optimal strategy for isolating MSCs from coagulated human bone marrow.
  • This method improves MSC recovery from challenging, poorly aspirated samples.
  • The technique facilitates the use of bone marrow samples that would otherwise be compromised.