Springer Protocols: Abstract: A Method to Isolate and Purify Human Bone Marrow Stromal Stem Cells

A Method to Isolate and Purify Human Bone Marrow Stromal Stem Cells

By: Stan Gronthos³, Andrew C. W. Zannettino³

Abstract

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The STRO-1 antibody can be used as a single reagent to isolate human bone marrow stromal stem cells (BMSSC), owing to its restricted specificity to a cell surface molecule expressed by clonogenic BMSSC, with little or no reactivity to hematopoietic stem/progenitor populations or mature stromal elements. The present protocol uses a combination of two different immunoselection methodologies in an attempt to generate highly purified preparations of BMSSC. This process involves the initial isolation of a minor subpopulation of bone marrow mononuclear cells (approx 10%) expressing the STRO-1 antigen, by means of magnetic activated cell sorting. Dual-color fluorescence activated cell sorting is then used as a secondary step to further purify the rare STRO-1^bright expressing fraction that contains all of the colony-forming BMSSC, based on their co-expression of a secondary cell surface marker, CD106 (VCAM-1).

Affiliation(s): (3) Mesenchymal Stem Cell Group, Division of Haematology, Institute of Medical and Veterinary Science, Hanson Institute, University of Adelaide, SA, Australia

Book Title: Mesenchymal Stem Cells: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 449 | Pub. Date: Apr-01-2008 | Page Range: 45-57 | DOI: 10.1007/978-1-60327-169-1_3

Subject: Cell Biology

Key Words: Bone marrow stromal stem cells - mesenchymal stem cells - magnetic activated cell sorting - fluorescence activated cell sorting

References

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