A Method to Isolate and Purify Human Bone Marrow Stromal Stem Cells
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-1bright expressing fraction that contains all of the colony-forming BMSSC, based on their co-expression of a secondary cell surface marker, CD106 (VCAM-1).
KeywordsBone marrow stromal stem cells mesenchymal stem cells magnetic activated cell sorting fluorescence activated cell sorting
The authors wish to acknowledge the technical assistance of Ms. Fiona Kohr and Ms. Sharon Paton. This work was supported by National Health and Medical Council of Australia grants (A Zannettino, S Gronthos).
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