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  • Original Article
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Pre-Clinical Studies

Isolation of mesenchymal stem cells from G-CSF-mobilized human peripheral blood using fibrin microbeads

Bone Marrow Transplantation volume 37, pages 967–976 (2006)Cite this article

Abstract

Adult mesenchymal stem cells (MSC) that are able to differentiate into various mesenchymal cell types are typically isolated from bone marrow, but their significant presence in human peripheral blood (PB) is controversial. Fibrin microbeads (FMB) that bind matrix-dependent cells were used to isolate MSC from the mononuclear fraction of mobilized PB of adult healthy human donors treated with a granulocyte colony-stimulating factor. Isolation by plastic adherence resulted in a negligible number of MSC in all samples tested, whereas FMB-based isolation yielded spindle-shaped cell samples that could further expand on plastic or on FMB in eight out of the 11 samples. The yield of these cells at days 17–18 after the harvest was ∼0.5% of the initial cell number. The isolated cells were grown on plastic and characterized by FACS analysis and immunohistochemistry for specific markers. Following culturing and first passage, the FMB-isolated cells stained positive for mesenchymal stromal cell markers CD90 and CD105, expressed vimentin and fibronectin and were negative for hematopoietic markers CD45 and CD34. These cells could differentiate into osteoblasts, adipocytes and chondrocytes. This study indicates that FMB may have special advantage in isolating MSC from sources such as mobilized PB, where the number of such cells is scarce.

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Acknowledgements

We thank Anna Hotovely-Solomon, Irena Shimeliovich and Elena Gaberman for their technical help and advice. This work was partially supported by the Israel Science Foundation Grant #697/001 to RG and by HAPTO Biotech Ltd.

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Authors and Affiliations

  1. Radiobiology and Biotechnology Laboratory, Sharett Institute of Oncology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    I Kassis, L Zangi, R Rivkin, L Levdansky & R Gorodetsky

  2. Department of Bone Marrow Transplantation, Hadassah-Hebrew University Medical Center, Jerusalem, Israel

    S Samuel

  3. HAPTO Biotech Ltd, Jerusalem, Israel

    G Marx & R Gorodetsky

Authors
  1. I Kassis
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  2. L Zangi
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  3. R Rivkin
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  4. L Levdansky
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  5. S Samuel
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  6. G Marx
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  7. R Gorodetsky
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Correspondence to R Gorodetsky.

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Kassis, I., Zangi, L., Rivkin, R. et al. Isolation of mesenchymal stem cells from G-CSF-mobilized human peripheral blood using fibrin microbeads. Bone Marrow Transplant 37, 967–976 (2006). https://doi.org/10.1038/sj.bmt.1705358

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